Ester and carbonate pyrimidine compounds as JAK kinase inhibitors

ABSTRACT

The invention provides compounds of formula (I): 
                         
or pharmaceutically-acceptable salts thereof, that are inhibitors of Janus kinases and release an active metabolite in vivo. The invention also provides pharmaceutical compositions comprising such compounds, and methods of using such compounds to treat inflammatory skin diseases and other diseases.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/837,829, filed on Apr. 24, 2019, the disclosure of which isincorporated herein by reference in its entirety.

BACKGROUND OF THE INVENTION Field of the Invention

The invention is directed to ester and carbonate pyrimidine compoundsuseful as JAK inhibitors which also release an active metabolite invivo. The invention is also directed to pharmaceutical compositionscomprising such compound, and methods of using such compounds to treatinflammatory and autoimmune diseases.

State of the Art

Inhibition of the family of JAK enzymes inhibits signaling of many keypro-inflammatory cytokines. Thus, JAK inhibitors are expected to beuseful in the treatment of atopic dermatitis and other inflammatory skindiseases. Atopic dermatitis (AD) is a common chronic inflammatory skindisease that affects an estimated 14 million people in the United Statesalone. It is estimated that AD affects 10 to 20% of children and 1 to 3%of adults in developed countries (Bao et al., JAK-STAT, 2013, 2, e24137)and the prevalence is increasing. Elevation of proinflammatory cytokinesthat rely on the JAK-STAT pathway, in particular, IL-4, IL-5, IL-10,IL-12, IL-13, IFNγ, and TSLP has been associated with AD (Bao et al.,Leung et al., The Journal of Clinical Investigation, 2004, 113,651-657). In addition, upregulation of IL-31, another cytokine thatsignals through a JAK pairing, has been shown to have a role in thepruritus associated with the chronic state of AD (Sonkoly et al.,Journal of Allergy and Clinical Immunology, 2006, 117, 411-417).

Due to the modulating effect of the JAK/STAT pathway on the immunesystem, systemic exposure to JAK inhibitors may have an adverse systemicimmunosuppressive effect. Therefore, it would be desirable to providenew JAK inhibitors which have their effect at the site of action withoutsignificant systemic effects. In particular, for the treatment ofinflammatory skin diseases, such as atopic dermatitis, it would bedesirable to provide new JAK inhibitors which can be administeredtopically and achieve therapeutically relevant exposure in the skin.There remains a need for JAK inhibitor compounds with adequatesolubility in aqueous and/or organic excipients allowing development offormulations for topical application.

SUMMARY OF THE INVENTION

In one aspect, the invention provides compounds having activity as a JAKinhibitors that also release an active metabolite in vivo.

Accordingly, the invention provides a compound of formula (I):

or a pharmaceutically-acceptable salt thereof,

wherein X is —O— or a bond;

R is selected from the group consisting of a C₁₋₈ alkyl, a 4 to 7membered heterocyclic group, and a 3 to 8 membered cycloalkyl group,wherein the C₁₋₈ alkyl, heterocyclic and cycloalkyl groups areoptionally substituted with 1 to 3 R^(a); wherein each R^(a) isindependently selected from the group consisting of C₁₋₄ alkyl, CN, F,OH, C₁₋₄ alkyl-OH, and C₁₋₄ alkoxy.

The invention also provides a pharmaceutical composition comprising acompound of formula (I), or a pharmaceutically-acceptable salt thereofand a pharmaceutically-acceptable carrier.

The invention also provides a compound of formula (I) or apharmaceutically-acceptable salt thereof for use as a medicament.

The invention also provides a compound of formula (I) or apharmaceutically-acceptable salt thereof as described herein for use intreating inflammatory and autoimmune diseases or disorders.

The invention also provides a compound of formula (I) or apharmaceutically-acceptable salt thereof for use in treating a diseasein a mammal for which a JAK inhibitor is indicated.

The invention also provides a method of treating a disease in a mammalfor which a JAK inhibitor is indicated, the method comprisingadministering a compound of formula (I), or a pharmaceuticallyacceptable salt thereof, to the mammal.

The invention also provides a method of treating inflammatory andautoimmune diseases of the skin in a mammal, in particular atopicdermatitis and alopecia areata, the method comprising administeringcompound (I), or a pharmaceutically acceptable salt thereof, to themammal.

DETAILED DESCRIPTION OF THE INVENTION

Among other aspects, the invention provides compounds of formula (I),and pharmaceutically-acceptable salts thereof, having activity as JAKinhibitors which also release an active metabolite.

The invention provides a compound of formula (I):

or a pharmaceutically-acceptable salt thereof,

wherein X is —O— or a bond;

R is selected from the group consisting of a C₁₋₈ alkyl, a 4 to 7membered heterocyclic group, and a 3 to 8 membered cycloalkyl group,wherein the C₁₋₈ alkyl, heterocyclic and cycloalkyl groups areoptionally substituted with 1 to 3 R^(a);

each R^(a) is independently selected from the group consisting of C₁₋₄alkyl, CN, F, OH, C₁₋₄ alkyl-OH, and C₁₋₄ alkoxy.

In some embodiments, R is selected from the group consisting of a C₁₋₆alkyl, a 5 to 7 membered heterocyclic group, and a 5 to 7 memberedcycloalkyl group, wherein the C₁₋₆ alkyl, heterocyclic and cycloalkylgroups are optionally substituted with 1 to 3 R^(a).

In some embodiments, R is selected from the group consisting of a C₁₋₆alkyl, a 5 to 6 membered heterocyclic group, and a 5 to 6 memberedcycloalkyl group, wherein the C₁₋₆ alkyl, heterocyclic and cycloalkylgroups are optionally substituted with 1 to 3 R^(a).

In some embodiments, R is selected from the group consisting of C₁₋₆alkyl, cyclohexyl, and tetrahydropyran, wherein the C₁₋₆ alkyl,cyclohexyl, and tetrahydropyran are optionally substituted with 1 to 2R^(a).

In some embodiments, R is selected from the group consisting ofunsubstituted C₁₋₆ alkyl, unsubstituted cyclohexyl, and unsubstitutedtetrahydropyran.

In some embodiments, R is selected from the group consisting of methyl,ethyl, isopropyl, propyl, n-butyl, n-hexyl, cyclohexyl, andtetrahydropyran.

In some embodiments, R is C₁₋₈ alkyl optionally substituted with 1 to 3R^(a).

In some embodiments, R is a 5 to 7 membered heterocyclic groupoptionally substituted with 1 to 3 R^(a).

In some embodiments, R is a 5 to 7 membered cycloalkyl group optionallysubstituted with 1 to 3 R^(a).

In some embodiments, X is a bond. In some embodiments, X is —O—.

In some embodiments, X is a bond and R is selected from the groupconsisting of a C₁₋₈ alkyl and a 4 to 7 membered heterocyclic group,wherein the C₁₋₈ alkyl and heterocyclic groups are optionallysubstituted with 1 to 3 R^(a), wherein each R^(a) is independentlyselected from the group consisting of C₁₋₄ alkyl, CN, F, OH, C₁₋₄alkyl-OH, and C₁₋₄ alkoxy.

In some embodiments, X is a bond and R is selected from the groupconsisting of C₁₋₆ alkyl and tetrahydropyran, wherein the C₁₋₆ alkyl andtetrahydropyran are optionally substituted with 1 to 2 R^(a).

In some embodiments, X is a bond and R is selected from the groupconsisting of methyl, ethyl, propyl, n-butyl, n-hexyl, andtetrahydropyran.

In some embodiments, X is —O— and R is selected from the groupconsisting of a C₁₋₆ alkyl, a 4 to 7 membered heterocyclic group, and a3 to 8 membered cycloalkyl group, wherein the C₁₋₆ alkyl, heterocyclicand cycloalkyl groups are optionally substituted with 1 to 3 R^(a),wherein each R^(a) is independently selected from the group consistingof C₁₋₄ alkyl, CN, F, OH, C₁₋₄ alkyl-OH, and C₁₋₄ alkoxy.

In some embodiments, X is —O— and R is selected from the groupconsisting of a 4 to 7 membered heterocyclic group and a 3 to 8 memberedcycloalkyl group, wherein the heterocyclic and cycloalkyl groups areoptionally substituted with 1 to 3 R^(a), wherein each R^(a) isindependently selected from the group consisting of C₁₋₄ alkyl, CN, F,OH, C₁₋₄ alkyl-OH, and C₁₋₄ alkoxy.

In some embodiments, X is —O— and R is selected from the groupconsisting of a 5 to 6 membered heterocyclic group, and a 5 to 6membered cycloalkyl group, wherein the heterocyclic and cycloalkylgroups are optionally substituted with 1 to 3 R^(a).

In some embodiments, X is —O— and R is selected from the groupconsisting of isopropyl, cyclohexyl, and tetrahydropyran.

The invention also provides a compound selected from the groupconsisting of:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is a compound of formula:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound is a compound of formula:

or a pharmaceutically acceptable salt thereof.

In some embodiments, the invention provides a pharmaceutical compositioncomprising a compound of the disclosure, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically-acceptable carrier. Insome embodiments, the pharmaceutical composition further comprises oneor more additional therapeutic agents. In some embodiments, thepharmaceutical composition is an ointment or a cream.

Chemical structures are named herein according to IUPAC conventions asimplemented in ChemDraw software (PerkinElmer, Inc., Cambridge, Mass.).For example, compound 1:

is designated as(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methylacetate. The (1R,3s,5S) notation describes the exo orientation of thepyrimidinylamino group with respect to the 9-azabicyclo[3.3.1]nonanegroup.

Furthermore, the pyrazolyl moiety of compound (I) as well as othercompounds disclosed herein exists in tautomeric form. It will beunderstood that although specific structures are shown, or named, in aparticular form, the invention also includes the tautomer thereof.

The compounds of the disclosure contain one or more chiral centers andtherefore, such compounds (and intermediates thereof) can exist asracemic mixtures; pure stereoisomers (i.e., enantiomers ordiastereomers); stereoisomer-enriched mixtures and the like. Chiralcompounds shown or named herein without a defined stereochemistry at achiral center are intended to include any or all possible stereoisomervariations at the undefined stereocenter unless otherwise indicated. Thedepiction or naming of a particular stereoisomer means the indicatedstereocenter has the designated stereochemistry with the understandingthat minor amounts of other stereoisomers may also be present unlessotherwise indicated, provided that the utility of the depicted or namedcompound is not eliminated by the presence of another stereoisomer.

The compounds of formula (I) may exist as a free form or in various saltforms, such a mono-protonated salt form, a di-protonated salt form, atri-protonated salt form, or mixtures thereof. All such forms areincluded within the scope of this invention, unless otherwise indicated.

This disclosure also includes isotopically-labeled versions of thecompounds of the disclosure, including compounds of formula (I), wherean atom has been replaced or enriched with an atom having the sameatomic number but an atomic mass different from the atomic mass thatpredominates in nature. Examples of isotopes that may be incorporatedinto a compound of formula (I) include, but are not limited to, ²H, ³H,¹¹C, ¹³C, ¹⁴C, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³⁵S, and ¹⁸F. Of particular interestare compounds of formula (I) enriched in tritium or carbon-14, whichcompounds can be used, for example, in tissue distribution studies. Alsoof particular interest are compounds of formula (I) enriched indeuterium especially at a site of metabolism, which compounds areexpected to have greater metabolic stability. Additionally of particularinterest are compounds of formula (I) enriched in a positron emittingisotope, such as ¹¹C, ¹⁸F, ¹⁵O and ¹³N, which compounds can be used, forexample, in Positron Emission Tomography (PET) studies.

Definitions

When describing this invention including its various aspects andembodiments, the following terms have the following meanings, unlessotherwise indicated.

The term “alkyl” means a monovalent saturated hydrocarbon group whichmay be linear or branched or combinations thereof. Unless otherwisedefined, such alkyl groups typically contain from 1 to 10 carbon atoms.Representative alkyl groups include, by way of example, methyl (Me),ethyl (Et), n-propyl (n-Pr) or (nPr), isopropyl (i-Pr) or (iPr), n-butyl(n-Bu) or (nBu), sec-butyl, isobutyl, tert-butyl (t-Bu) or (tBu),n-pentyl, n-hexyl, 2,2-dimethylpropyl, 2-methylbutyl, 3-methylbutyl,2-ethylbutyl, 2,2-dimethylpentyl, 2-propylpentyl, and the like.

When a specific number of carbon atoms are intended for a particularterm, the number of carbon atoms is shown preceding the term. Forexample, the term “C₁₋₃alkyl” means an alkyl group having from 1 to 3carbon atoms wherein the carbon atoms are in any chemically-acceptableconfiguration, including linear or branched configurations.

The term “alkoxy” means the monovalent group —O-alkyl, where alkyl isdefined as above. Representative alkoxy groups include, by way ofexample, methoxy, ethoxy, propoxy, butoxy, and the like.

The term “cycloalkyl” means a monovalent saturated carbocyclic groupwhich may be monocyclic or multicyclic. Unless otherwise defined, suchcycloalkyl groups typically contain from 3 to 10 carbon atoms.Representative cycloalkyl groups include, by way of example, cyclopropyl(cPr), cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl,adamantyl, and the like.

The term “halogen” means fluoro, chloro, bromo or iodo.

The term “heterocyclyl”, “heterocycle”, “heterocyclic”, or “heterocyclicring” means a monovalent saturated or partially unsaturated cyclicnon-aromatic group, having from 3 to 10 total ring atoms, wherein thering contains from 2 to 9 carbon ring atoms and from 1 to 4 ringheteroatoms selected from nitrogen, oxygen, and sulfur. Heterocyclicgroups may be monocyclic or multicyclic (i.e., fused or bridged).Representative heterocyclic groups include, by way of example,pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, morpholinyl,thiomorpholyl, indolin-3-yl, 2-imidazolinyl, tetrahydropyranyl,1,2,3,4-tetrahydroisoquinolin-2-yl, quinuclidinyl, 7-azanorbornanyl,nortropanyl, and the like, where the point of attachment is at anyavailable carbon or nitrogen ring atom. Where the context makes thepoint of attachment of the heterocyclic group evident, such groups mayalternatively be referred to as a non-valent species, i.e. pyrrolidine,piperidine, piperazine, imidazole, tetrahydropyran etc.

The term “therapeutically effective amount” means an amount sufficientto effect treatment when administered to a patient in need of treatment.

The term “treatment” as used herein means the treatment of a disease,disorder, or medical condition (such as a gastrointestinal inflammatorydisease), in a patient, such as a mammal (particularly a human) whichincludes one or more of the following:

(a) preventing the disease, disorder, or medical condition fromoccurring, i.e., preventing the reoccurrence of the disease or medicalcondition or prophylactic treatment of a patient that is pre-disposed tothe disease or medical condition;

(b) ameliorating the disease, disorder, or medical condition, i.e.,eliminating or causing regression of the disease, disorder, or medicalcondition in a patient, including counteracting the effects of othertherapeutic agents;

(c) suppressing the disease, disorder, or medical condition, i.e.,slowing or arresting the development of the disease, disorder, ormedical condition in a patient; or

(d) alleviating the symptoms of the disease, disorder, or medicalcondition in a patient.

The term “pharmaceutically acceptable salt” means a salt that isacceptable for administration to a patient or a mammal, such as a human(e.g., salts having acceptable mammalian safety for a given dosageregime). Representative pharmaceutically acceptable salts include saltsof acetic, ascorbic, benzenesulfonic, benzoic, camphorsulfonic, citric,ethanesulfonic, edisylic, fumaric, gentisic, gluconic, glucoronic,glutamic, hippuric, hydrobromic, hydrochloric, isethionic, lactic,lactobionic, maleic, malic, mandelic, methanesulfonic, mucic,naphthalenesulfonic, naphthalene-1,5-disulfonic,naphthalene-2,6-disulfonic, nicotinic, nitric, orotic, pamoic,pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonicand xinafoic acid, and the like.

The term “salt thereof” means a compound formed when the hydrogen of anacid is replaced by a cation, such as a metal cation or an organiccation and the like. For example, the cation can be a protonated form ofa compound of formula (I), i.e. a form where one or more amino groupshave been protonated by an acid. Typically, the salt is apharmaceutically acceptable salt, although this is not required forsalts of intermediate compounds that are not intended for administrationto a patient.

General Synthetic Procedures

Compounds of formula (I), and intermediates thereof, can be preparedaccording to the following general methods and procedures usingcommercially-available or routinely-prepared starting materials andreagents. The substituents and variables (e.g., R, X, R^(a) etc) used inthe following schemes have the same meanings as those defined elsewhereherein unless otherwise indicated. Additionally, compounds having anacidic or basic atom or functional group may be used or may be producedas a salt unless otherwise indicated (in some cases, the use of a saltin a particular reaction will require conversion of the salt to anon-salt form, e.g., a free base, using routine procedures beforeconducting the reaction).

Although a particular embodiment of the present invention may be shownor described in the following procedures, those skilled in the art willrecognize that other embodiments or aspects of the present invention canalso be prepared using such procedures or by using other methods,reagents, and starting materials known to those skilled in the art. Inparticular, it will be appreciated that compounds of formula (I) may beprepared by a variety of process routes in which reactants are combinedin different orders to provide different intermediates en route toproducing the final product.

General methods of preparing compounds of formula (I) are illustrated inscheme 1.

Compound M may be converted to a compound of formula (I) by reactionwith S-1 in the presence of a base, where LG is a leaving group. In someembodiments, the leaving group is chloro and S-1 is either an acylchloride or a chloroformate. In some embodiments, X is a bond and thereaction is conducted in the presence of DIPEA and DMAP. In someembodiments, X is O and the reaction is conducted in the presence ofpyridine and either DMF or THF.

Alternatively, when X is a bond, compound M may be converted to acompound of formula (I) by coupling with a carboxylic acid S-2 in thepresence of a coupling agent such as HATU. In some embodiments, thereaction is conducted in presence of DIPEA and DMAP.

Pharmaceutical Compositions

Compounds of formula (I) and pharmaceutically-acceptable salts thereofare typically used in the form of a pharmaceutical composition orformulation. Such pharmaceutical compositions may be administered to apatient by any acceptable route of administration including, but notlimited to, oral, topical (including transdermal), rectal, nasal,inhaled, and parenteral modes of administration.

Accordingly, in one of its composition aspects, the invention isdirected to a pharmaceutical composition comprising apharmaceutically-acceptable carrier or excipient and a compound offormula (I), or a pharmaceutically-acceptable salt thereof. Optionally,such pharmaceutical compositions may contain other therapeutic and/orformulating agents if desired. When discussing compositions and usesthereof, the “compound of the invention” may also be referred to hereinas the “active agent”.

The pharmaceutical compositions of this disclosure typically contain atherapeutically effective amount of a compound of formula (I), or apharmaceutically-acceptable salt thereof. Those skilled in the art willrecognize, however, that a pharmaceutical composition may contain morethan a therapeutically effective amount, i.e., bulk compositions, orless than a therapeutically effective amount, i.e., individual unitdoses designed for multiple administration to achieve a therapeuticallyeffective amount.

Typically, such pharmaceutical compositions will contain from about 0.1to about 95% by weight of the active agent; including from about 5 toabout 70% by weight of the active agent.

Any conventional carrier or excipient may be used in the pharmaceuticalcompositions of the invention. The choice of a particular carrier orexcipient, or combinations of carriers or excipients, will depend on themode of administration being used to treat a particular patient or typeof medical condition or disease state. In this regard, the preparationof a suitable pharmaceutical composition for a particular mode ofadministration is well within the scope of those skilled in thepharmaceutical arts. Additionally, the carriers or excipients used inthe pharmaceutical compositions of this invention arecommercially-available. By way of further illustration, conventionalformulation techniques are described in Remington: The Science andPractice of Pharmacy, 20th Edition, Lippincott Williams & White,Baltimore, Md. (2000); and H.C. Ansel et al., Pharmaceutical DosageForms and Drug Delivery Systems, 7th Edition, Lippincott Williams &White, Baltimore, Md. (1999).

Representative examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, the following:sugars, such as lactose, glucose and sucrose; starches, such as cornstarch and potato starch; cellulose, such as microcrystalline cellulose,and its derivatives, such as sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin;talc; excipients, such as cocoa butter and suppository waxes; oils, suchas peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil,corn oil and soybean oil; glycols, such as propylene glycol; polyols,such as glycerin, sorbitol, mannitol and polyethylene glycol; esters,such as ethyl oleate and ethyl laurate; agar; buffering agents, such asmagnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-freewater; isotonic saline; Ringer's solution; ethyl alcohol; phosphatebuffer solutions; and other non-toxic compatible substances employed inpharmaceutical compositions.

Pharmaceutical compositions are typically prepared by thoroughly andintimately mixing or blending the active agent with apharmaceutically-acceptable carrier and one or more optionalingredients. The resulting uniformly blended mixture may then be shapedor loaded into tablets, capsules, pills and the like using conventionalprocedures and equipment.

The pharmaceutical compositions of this disclosure may be packaged in aunit dosage form. The term “unit dosage form” refers to a physicallydiscrete unit suitable for dosing a patient, i.e., each unit containinga predetermined quantity of active agent calculated to produce thedesired therapeutic effect either alone or in combination with one ormore additional units. For example, such unit dosage forms may becapsules, tablets, pills, and the like, or unit packages suitable forparenteral administration.

In one embodiment, the pharmaceutical compositions of the invention aresuitable for oral administration. Suitable pharmaceutical compositionsfor oral administration may be in the form of capsules, tablets, pills,lozenges, cachets, dragees, powders, granules; or as a solution or asuspension in an aqueous or non-aqueous liquid; or as an oil-in-water orwater-in-oil liquid emulsion; or as an elixir or syrup; and the like;each containing a predetermined amount of a compound of this disclosure,or a pharmaceutically-acceptable salt thereof, as an active ingredient.

When intended for oral administration in a solid dosage form (i.e., ascapsules, tablets, pills and the like), the pharmaceutical compositionsof this disclosure will typically comprise the active agent, or apharmaceutically acceptable salt thereof, and one or morepharmaceutically-acceptable carriers. Optionally, such solid dosageforms may comprise: fillers or extenders, such as starches,microcrystalline cellulose, lactose, dicalcium phosphate, sucrose,glucose, mannitol, and/or silicic acid; binders, such ascarboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone,sucrose and/or acacia; humectants, such as glycerol; disintegratingagents, such as crosscarmellose sodium, agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, certain silicates, and/or sodiumcarbonate; solution retarding agents, such as paraffin; absorptionaccelerators, such as quaternary ammonium compounds; wetting agents,such as cetyl alcohol and/or glycerol monostearate; absorbents, such askaolin and/or bentonite clay; lubricants, such as talc, calciumstearate, magnesium stearate, solid polyethylene glycols, sodium laurylsulfate, and/or mixtures thereof; coloring agents; and buffering agents.

Release agents, wetting agents, coating agents, sweetening, flavoringand perfuming agents, preservatives and antioxidants can also be presentin the pharmaceutical compositions of this disclosure. Examples ofpharmaceutically-acceptable antioxidants include: water-solubleantioxidants, such as ascorbic acid, cysteine hydrochloride, sodiumbisulfate, sodium metabisulfate, sodium sulfite and the like;oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole, butylated hydroxytoluene, lecithin, propyl gallate,alpha-tocopherol, and the like; and metal-chelating agents, such ascitric acid, ethylenediamine tetraacetic acid, sorbitol, tartaric acid,phosphoric acid, and the like. Coating agents for tablets, capsules,pills and like, include those used for enteric coatings, such ascellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropylmethylcellulose phthalate, methacrylic acid, methacrylic acid estercopolymers, cellulose acetate trimellitate, carboxymethyl ethylcellulose, hydroxypropyl methyl cellulose acetate succinate, and thelike.

Pharmaceutical compositions of this disclosure may also be formulated toprovide slow or controlled release of the active agent using, by way ofexample, hydroxypropyl methylcellulose in varying proportions; or otherpolymer matrices, liposomes and/or microspheres. In addition, thepharmaceutical compositions of this disclosure may optionally containopacifying agents and may be formulated so that they release the activeingredient only, or preferentially, in a certain portion of thegastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions which can be used include polymeric substancesand waxes. The active agent can also be in micro-encapsulated form, ifappropriate, with one or more of the above-described excipients.

Suitable liquid dosage forms for oral administration include, by way ofillustration, pharmaceutically-acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. Liquid dosage formstypically comprise the active agent and an inert diluent, such as, forexample, water or other solvents, solubilizing agents and emulsifiers,such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, oils (esp., cottonseed, groundnut, corn, germ, olive, castor andsesame oils), oleic acid, glycerol, tetrahydrofuryl alcohol,polyethylene glycols and fatty acid esters of sorbitan, and mixturesthereof. Alternatively, certain liquid formulations can be converted,for example, by spray drying, to a powder, which is used to preparesolid dosage forms by conventional procedures.

Suspensions, in addition to the active ingredient, or a pharmaceuticallyacceptable salt thereof, may contain suspending agents such as, forexample, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol andsorbitan esters, microcrystalline cellulose, aluminum metahydroxide,bentonite, agar-agar and tragacanth, and mixtures thereof.

A compound of formula (I), or a pharmaceutically-acceptable saltthereof, may also be administered parenterally (e.g. by intravenous,subcutaneous, intramuscular or intraperitoneal injection). Forparenteral administration, the active agent, or a pharmaceuticallyacceptable salt thereof, is typically admixed with a suitable vehiclefor parenteral administration including, by way of example, sterileaqueous solutions, saline, low molecular weight alcohols such aspropylene glycol, polyethylene glycol, vegetable oils, gelatin, fattyacid esters such as ethyl oleate, and the like. Parenteral formulationsmay also contain one or more anti-oxidants, solubilizers, stabilizers,preservatives, wetting agents, emulsifiers, buffering agents, ordispersing agents. These formulations may be rendered sterile by use ofa sterile injectable medium, a sterilizing agent, filtration,irradiation, or heat.

Alternatively, the pharmaceutical compositions of this disclosure areformulated for administration by inhalation. Suitable pharmaceuticalcompositions for administration by inhalation will typically be in theform of an aerosol or a powder. Such compositions are generallyadministered using well-known delivery devices, such as a metered-doseinhaler, a dry powder inhaler, a nebulizer or a similar delivery device.

When administered by inhalation using a pressurized container, thepharmaceutical compositions of this disclosure will typically comprisethe active ingredient, or a pharmaceutically acceptable salt thereof,and a suitable propellant, such as dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. Additionally, the pharmaceutical composition may bein the form of a capsule or cartridge (made, for example, from gelatin)comprising a compound of the disclosure, or apharmaceutically-acceptable salt thereof, and a powder suitable for usein a powder inhaler. Suitable powder bases include, by way of example,lactose or starch.

Topical Formulations

To treat skin conditions, the compound of the disclosure, or apharmaceutically-acceptable salt thereof, is preferably formulated fortopical administration to the skin. Topical compositions comprise fluidor semi-solid vehicles that may include but are not limited to polymers,thickeners, buffers, neutralizers, chelating agents, preservatives,surfactants or emulsifiers, antioxidants, waxes or oils, emollients,sunscreens, and a solvent or mixed solvent system. The topicalcompositions useful in the subject invention can be made into a widevariety of product types. These include, but are not limited to lotions,creams, gels, sticks, sprays, ointments, pastes, foams, mousses, andcleansers. These product types can comprise several types of carriersystems including, but not limited to particles, nanoparticles, andliposomes. If desired, disintegrating agents can be added, such as thecross-linked polyvinyl pyrrolidone, agar or alginic acid or a saltthereof such as sodium alginate. Techniques for formulation andadministration can be found in Remington: The Science and Practice ofPharmacy, 19th Ed. (Easton, Pa.: Mack Publishing Co., 1995). Theformulation can be selected to maximize delivery to a desired targetsite in the body.

Lotions, which are preparations that are to be applied to the skin, orhair surface without friction, are typically liquid or semi-liquidpreparations in which finely divided solid, waxy, or liquid aredispersed. Lotions will typically contain suspending agents to producebetter dispersions as well as compounds useful for localizing andholding the active agent in contact with the skin or hair, e.g.,methylcellulose, sodium carboxymethyl-cellulose, or the like.

Creams containing the active agent, or a pharmaceutically acceptablesalt thereof, for delivery according to the present disclosure areviscous liquid or semisolid emulsions, either oil-in-water orwater-in-oil. Cream bases are water-washable, and contain an oil phase,an emulsifier and an aqueous phase. The oil phase is generally comprisedof petrolatum or a fatty alcohol, such as cetyl- or stearyl alcohol; theaqueous phase usually, although not necessarily, exceeds the oil phasein volume, and generally contains a humectant. The emulsifier in a creamformulation, as explained in Remington: The Science and Practice ofPharmacy, is generally a nonionic, anionic, cationic or amphotericsurfactant. Components of cream formulations may include: oil bases,such as petrolatrum, mineral oils, vegetable and animal oils, andtriglycerides; cream bases, such as lanolin alcohols, stearic acid, andcetostearyl alcohol; a gel base, such as polyvinyl alcohol; solvents,such as, propylene glycol and polyethylene glycol; emulsifiers, such aspolysorbates, stearates, such as glyceryl stearate,octylhydroxystearate, polyoxyl stearate, PEG stearyl ethers, isopropylpalmitate, and sorbitan monostearate; stabilizers, such aspolysaccharides and sodium sulfite; emollients (i.e. moisturizers), suchas medium chain triglycerides, isopropyl myristate, and dimethicone;stiffening agents, such as cetyl alcohol and stearyl alcohol;antimicrobial agents, such as methylparaben, propylparaben,phenoxyethanol, sorbic acid, diazolidinyl urea, and butylatedhydroxyanisole; penetration enhancers, such as N-methylpyrrolidone,propylene glycol, polyethylene glycol monolaurate, and the like; andchelating agents, such as edetate disodium.

Gel formulations can also be used in connection with the presentinvention. As will be appreciated by those working in the field oftopical drug formulation, gels are semisolid. Single-phase gels containorganic macromolecules distributed substantially uniformly throughoutthe carrier liquid, which is typically aqueous, but also may be asolvent or solvent blend.

Ointments, which are semisolid preparations, are typically based onpetrolatum or other petroleum derivatives. As will be appreciated by theordinarily skilled artisan, the specific ointment base to be used is onethat provides for optimum delivery for the active agent chosen for agiven formulation, and, preferably, provides for other desiredcharacteristics as well, e.g., emolliency or the like. As with othercarriers or vehicles, an ointment base should be inert, stable,nonirritating and non-sensitizing. As explained in Remington: TheScience and Practice of Pharmacy, 19th Ed. (Easton, Pa.: Mack PublishingCo., 1995), at pages 1399-1404, ointment bases may be grouped in fourclasses: oleaginous bases; emulsifiable bases; emulsion bases; andwater-soluble bases. Oleaginous ointment bases include, for example,vegetable oils, fats obtained from animals, and semisolid hydrocarbonsobtained from petroleum. Emulsifiable ointment bases, also known asabsorbent ointment bases, contain little or no water and include, forexample, hydroxystearin sulfate, anhydrous lanolin and hydrophilicpetrolatum. Emulsion ointment bases are either water-in-oil (W/O)emulsions or oil-in-water (O/W) emulsions, and include, for example,cetyl alcohol, glyceryl monostearate, lanolin and stearic acid.Water-soluble ointment bases may be prepared from polyethylene glycolsof varying molecular weight; again, reference may be had to Remington:The Science and Practice of Pharmacy, supra, for further information.Suitable oily materials for use in ointment formulations includepetrolatum (petroleum jelly), beeswax, cocoa butter, shea butter, andcetyl alcohol. Ointments may optionally additionally include penetrationenhancers, if desired.

Useful formulations of this disclosure also encompass sprays. Spraysgenerally provide the active agent in an aqueous and/or alcoholicsolution which can be misted onto the skin or hair for delivery. Suchsprays include those formulated to provide for concentration of theactive agent solution at the site of administration following delivery,e.g., the spray solution can be primarily composed of alcohol or otherlike volatile liquid in which the drug or active agent can be dissolved.Upon delivery to the skin or hair, the carrier evaporates, leavingconcentrated active agent at the site of administration.

The topical pharmaceutical compositions may also comprise suitable solidor gel phase carriers. Examples of such carriers include but are notlimited to calcium carbonate, calcium phosphate, various sugars,starches, cellulose derivatives, gelatin, and polymers such aspolyethylene glycols.

The topical pharmaceutical compositions may also comprise a suitableemulsifier which refers to an agent that enhances or facilitates mixingand suspending oil-in-water or water-in-oil. The emulsifying agent usedherein may consist of a single emulsifying agent or may be a nonionic,anionic, cationic or amphoteric surfactant or blend of two or more suchsurfactants; preferred for use herein are nonionic or anionicemulsifiers. Such surface-active agents are described in “McCutcheon'sDetergent and Emulsifiers,” North American Edition, 1980 Annualpublished by the McCutcheon Division, MC Publishing Company, 175 RockRoad, Glen Rock, N.J. 07452, USA.

High molecular weight alcohols may be used such as cetearyl alcohol,cetyl alcohol, stearyl alcohol, emulsifying wax, glyceryl monostearate.Other examples are ethylene glycol distearate, sorbitan tristearate,propylene glycol monostearate, sorbitan monooleate, sorbitanmonostearate (SPAN 60), diethylene glycol monolaurate, sorbitanmonopalmitate, sucrose dioleate, sucrose stearate (CRODESTA F-160),polyoxyethylene lauryl ether (BRIJ 30), polyoxyethylene (2) stearylether (BRIJ 72), polyoxyethylene (21) stearyl ether (BRIJ 721),polyoxyethylene monostearate (Myrj 45), polyoxyethylene sorbitanmonostearate (TWEEN 60), polyoxyethylene sorbitan monooleate (TWEEN 80),polyoxyethylene sorbitan monolaurate (TWEEN 20) and sodium oleate.Cholesterol and cholesterol derivatives may also be employed inexternally used emulsions.

Example of suitable nonionic emulsifying agents are described by Paul L.Lindner in “Emulsions and Emulsion”, edited by Kenneth Lissant,published by Dekker, New York, N.Y., 1974. Examples of nonionicemulsifiers that may be used include but are not limited to BRIJproducts such as BRIJ 2 (a polyoxyethylene (2) stearyl ether), BRIJ S20(a polyoxyethylene (20) stearyl ether), BRIJ 72 (a polyoxyethylene (2)stearyl ether having an HLB of 4.9), BRIJ 721 (a polyoxyethylene (21)stearyl ether having an HLB of 15.5), Brij 30 (a polyoxyethylene laurylether having an HLB of 9.7), Polawax (emulsifying wax having an HLB of8.0), Span 60 (sorbitan monostearate having an HLB of 4.7), CrodestaF-160 (sucrose stearate” having an HLB of 14.5).

The topical pharmaceutical compositions may also comprise suitableemollients. Emollients are materials used for the prevention or reliefof dryness, as well as for the protection of the skin or hair. Usefulemollients include, but are not limited to, cetyl alcohol, isopropylmyristate, stearyl alcohol, and the like. A wide variety of suitableemollients are known and can be used herein. See e.g., Sagarin,Cosmetics, Science and Technology, 2nd Edition, Vol. 1, pp. 32-43(1972), and U.S. Pat. No. 4,919,934, to Deckner et al., issued Apr. 24,1990, both of which are incorporated herein by reference in theirentirety.

The topical pharmaceutical compositions may also comprise suitableantioxidants, substances known to inhibit oxidation. Antioxidantssuitable for use in accordance with the present invention include, butare not limited to, butylated hydroxytoluene, ascorbic acid, sodiumascorbate, calcium ascorbate, ascorbic palmitate, butylatedhydroxyanisole, 2,4,5-trihydroxybutyrophenone,4-hydroxymethyl-2,6-di-tert-butylphenol, erythorbic acid, gum guaiac,propyl gallate, thiodipropionic acid, dilauryl thiodipropionate,tert-butylhydroquinone and tocopherols such as vitamin E, and the like,including pharmaceutically acceptable salts and esters of thesecompounds. Preferably, the antioxidant is butylated hydroxytoluene,butylated hydroxyanisole, propyl gallate, ascorbic acid,pharmaceutically acceptable salts or esters thereof, or mixturesthereof. Most preferably, the antioxidant is butylated hydroxytoluene.

The topical pharmaceutical compositions may also comprise suitablepreservatives. Preservatives are compounds added to a pharmaceuticalformulation to act as an anti-microbial agent. Among preservatives knownin the art as being effective and acceptable in parenteral formulationsare benzalkonium chloride, benzethonium, chlorohexidine, phenol,m-cresol, benzyl alcohol, methylparaben, propylparaben, chlorobutanol,o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal,benzoic acid, and various mixtures thereof. See, e.g., Wallhausser, K.-H., Develop. Biol. Standard, 24:9-28 (1974) (S. Krager, Basel).

The topical pharmaceutical compositions may also comprise suitablechelating agents to form complexes with metal cations that do not crossa lipid bilayer. Examples of suitable chelating agents include ethylenediamine tetraacetic acid (EDTA), ethylene glycol-bis(beta-aminoethylether)-N,N,N′,N′-tetraacetic acid (EGTA) and8-amino-2-[(2-amino-5-methylphenoxy)methyl]-6-methoxyquinoline-N,N,N′,N′-tetraaceticacid, tetrapotassium salt (QUIN-2). Preferably the chelating agents areEDTA and citric acid.

The topical pharmaceutical compositions may also comprise suitableneutralizing agents used to adjust the pH of the formulation to within apharmaceutically acceptable range. Examples of neutralizing agentsinclude but are not limited to trolamine, tromethamine, sodiumhydroxide, hydrochloric acid, citric acid, and acetic acid.

The topical pharmaceutical compositions may also comprise suitableviscosity increasing agents. These components are diffusible compoundscapable of increasing the viscosity of a polymer-containing solutionthrough the interaction of the agent with the polymer. Carbopol Ultrez10 may be used as a viscosity-increasing agent.

Liquid forms, such as lotions suitable for topical administration mayinclude a suitable aqueous or non-aqueous vehicle with buffers,suspending and dispensing agents, thickeners, penetration enhancers, andthe like. Solid forms such as creams or pastes or the like may include,for example, any of the following ingredients, water, oil, alcohol orgrease as a substrate with surfactant, polymers such as polyethyleneglycol, thickeners, solids and the like. Liquid or solid formulationsmay include enhanced delivery technologies such as liposomes,microsomes, microsponges and the like. Additionally, the compounds canbe delivered using a sustained-release system, such as semipermeablematrices of solid hydrophobic polymers containing the therapeutic agent.Various sustained-release materials have been established and are wellknown by those skilled in the art.

When formulated for topical application, a compound of formula (I), or apharmaceutically-acceptable salt thereof, may be present at between 0.1and 50% by weight. In some embodiments, a compound of formula (I), or apharmaceutically-acceptable salt thereof, is present at between 0.1 and25% by weight. In some embodiments, a compound of formula (I), or apharmaceutically-acceptable salt thereof, is present at between 0.1 and10% by weight. In some embodiments, a compound of formula (I), or apharmaceutically-acceptable salt thereof, is present at between 0.25 and5% by weight. In some embodiments, a compound of formula (I), or apharmaceutically-acceptable salt thereof, is present at between 0.25 and2% by weight. In some embodiments, a compound of formula (I), or apharmaceutically-acceptable salt thereof, is present at between 0.25 and1% by weight. In some embodiments, a compound of formula (I), or apharmaceutically-acceptable salt thereof, is present at between 0.05 and0.5% by weight. In some embodiments, a compound of formula (I), or apharmaceutically-acceptable salt thereof, is present at about 0.1, 0.2,0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7,1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.25, 3.5,3.75, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5 or 10% by weight.

In some embodiments, the pharmaceutical composition comprising acompound of formula (I), or a pharmaceutically-acceptable salt thereof,further comprises one or more additional therapeutic agents. In someembodiments, the one or more additional therapeutic agents is useful totreat an autoimmune skin disease. In some embodiments, the one or moreadditional therapeutic agents is useful to treat an inflammatory skindisease. In some embodiments, the one or more additional therapeuticagents is useful to treat atopic dermatitis. In some embodiments, theone or more additional therapeutic agents is useful to treat alopeciaareata. Specific class of compounds or specific compounds that may becombined with a compound of formula (I) in a pharmaceutical compositionare exemplified in later paragraphs.

The following non-limiting examples illustrate representativepharmaceutical compositions of the present invention.

Tablet Oral Solid Dosage Form

A compound of formula (I) or a pharmaceutically-acceptable salt thereofis dry blended with microcrystalline cellulose, polyvinyl pyrrolidone,and croscarmellose sodium in a ratio of 4:5:1:1 and compressed intotablets to provide a unit dosage of, for example, 5 mg, 20 mg or 40 mgactive agent per tablet.

Capsule Oral Solid Dosage Form

A compound of formula (I) or a pharmaceutically-acceptable salt thereofis combined with microcrystalline cellulose, polyvinyl pyrrolidone, andcrosscarmellose sodium in a ratio of 4:5:1:1 by wet granulation andloaded into gelatin or hydroxypropyl methylcellulose capsules to providea unit dosage of, for example, 5 mg, 20 mg or 40 mg active agent percapsule.

Liquid Formulation

A liquid formulation comprising a compound of formula (I) or apharmaceutically-acceptable salt thereof (0.1%), water (98.9%) andascorbic acid (1.0%) is formed by adding a compound of the disclosure,or a pharmaceutically-acceptable salt thereof, to a mixture of water andascorbic acid.

Enteric Coated Oral Dosage Form

A compound of formula (I) or a pharmaceutically-acceptable salt thereof,is dissolved in an aqueous solution containing polyvinyl pyrrolidone andspray coated onto microcrystalline+cellulose or sugar beads in a ratioof 1:5 w/w active agent:beads and then an approximately 5% weight gainof an enteric coating comprising an acrylic copolymer, for example acombination of acrylic copolymers available under the trade namesEudragit-L® and Eudragit-S®, or hydroxypropyl methylcellulose acetatesuccinate is applied. The enteric coated beads are loaded into gelatinor hydroxypropyl methylcellulose capsules to provide a unit dosage of,for example, 30 mg active agent per capsule.

Enteric Coated Oral Dosage Form

An enteric coating comprising a combination of Eudragit-L® andEudragit-S®, or hydroxypropyl methylcellulose acetate succinate isapplied to a tablet oral dosage form or a capsule oral dosage formdescribed above.

Ointment Formulation for Topical Administration

A compound of formula (I) or a pharmaceutically-acceptable salt thereofis combined with petrolatum, C₈-C₁₀ triglyceride, octylhydroxystearate,and N-methylpyrrolidone in a ratio to provide a composition containing0.05% to 5% of active agent by weight.

Ointment Formulation for Topical Administration

A compound of formula (I) or a pharmaceutically-acceptable salt thereofis combined with petrolatum, C₈-C₁₀ triglyceride, octylhydroxystearate,benzyl alcohol and N-methylpyrrolidone in a ratio to provide acomposition containing 0.05% to 5% of active agent by weight.

Ointment Formulation for Topical Administration

A compound of formula (I) or a pharmaceutically-acceptable salt thereofis combined with white petrolatum, propylene glycol, mono- anddi-glycerides, paraffin, butylated hydroxytoluene, and edetate calciumdisodium in a ratio to provide a composition containing 0.05% to 5%active agent by weight.

Ointment Formulation for Topical Administration

A compound of formula (I) or a pharmaceutically-acceptable salt thereofis combined with mineral oil, paraffin, propylene carbonate, whitepetrolatum and white wax to provide a composition containing 0.05% to 5%active agent by weight.

Cream Formulation for Topical Administration

Mineral oil is combined with a compound of formula (I) or apharmaceutically-acceptable salt thereof, propylene glycol, isopropylpalmitate, polysorbate 60, cetyl alcohol, sorbitan monostearate,polyoxyl 40 stearate, sorbic acid, methylparaben and propylparaben toform an oil phase, which is combined with purified water by shearblending to provide a composition containing 0.05% to 5% active agent byweight.

Cream Formulation for Topical Administration

A cream formulation comprising a compound of formula (I) or apharmaceutically-acceptable salt thereof, benzyl alcohol, cetyl alcohol,citric acid anhydrous, mono and di-glycerides, oleyl alcohol, propyleneglycol, sodium cetostearyl sulphate, sodium hydroxide, stearyl alcohol,triglycerides, and water contains 0.05% to 5% active agent by weight.

Cream Formulation for Topical Administration

A cream formulation comprising a compound of formula (I) or apharmaceutically-acceptable salt thereof, cetostearyl alcohol, isopropylmyristate, propylene glycol, cetomacrogol 1000, dimethicone 360, citricacid, sodium citrate, and purified water, with imidurea, methylparaben,and propylparaben, as preservatives, contains 0.05% to 5% active agentby weight.

Cream Formulation for Topical Administration

A cream formulation comprising a compound of formula (I) or apharmaceutically-acceptable salt thereof, stearic acid, cetostearylalcohol, isopropyl palmitate, octylhydroxystearate, BRIJ S2 (PEG 2Stearyl Ether), BRIJ S20 (PEG 20 Stearyl Ether), N-Methylpyrrolidine,PEG and water contains 0.05% to 5% active agent by weight.

Cream Formulation for Topical Administration

A cream formulation comprising a compound of formula (I) or apharmaceutically-acceptable salt thereof, stearic acid, cetostearylalcohol, isopropyl palmitate, octylhydroxystearate, BRIJ S2 (PEG 2Stearyl Ether), BRIJ S20 (PEG 20 Stearyl Ether), N-Methylpyrrolidine,PEG400 and water contains 0.05% to 5% active agent by weight.

Utility

Compounds of formula (I) have been shown to be potent inhibitors of theJAK family of enzymes: JAK1, JAK2, JAK3, and TYK2. Additionally, theyhave the ability to release an active metabolite. Inhibition of thefamily of JAK enzymes inhibits signaling of many key pro-inflammatorycytokines. Thus compounds of formula (I) are expected to be useful inthe treatment of inflammatory diseases such as inflammatory and pruriticskin diseases, gastrointestinal inflammatory diseases inflammatoryocular diseases and inflammatory respiratory diseases.

Inflammatory Skin Disease

Atopic dermatitis has been associated with elevation of proinflammatorycytokines that rely on the JAK-STAT pathway, in particular, IL-4, IL-5,IL-10, IL-13, and IFNγ. Since compounds of formula (I) and their activemetabolite exhibit potent inhibition at all four JAK enzymes, they areexpected to potently inhibit the proinflammatory cytokinescharacteristic of atopic dermatitis and other inflammatory skindiseases. Compounds of formula (I) and their active metabolite were alsoshown here to exhibit high pIC₅₀ values for inhibition of IL-2 inducedSTATS phosphorylation in a cellular assay.

It is expected that sustained dermal levels of JAK inhibitors in theabsence of significant systemic levels will result in potent localanti-inflammatory and anti-pruritic activity in the skin withoutsystemically-driven adverse effects. Such compounds are expected to bebeneficial in a number of dermal inflammatory or pruritic conditionsthat include, but are not limited to atopic dermatitis, vitiligo,non-segmental vitiligo, cutaneous T cell lymphoma and subtypes (Sezarysyndrome, mycosis fungoides, pagetoid reticulosis, granulomatous slackskin, lymphomatoid papulosis, pityriasis lichenoides chronica,pityriasis lichenoides et varioliformis acuta, CD30+ cutaneous T-celllymphoma, secondary cutaneous CD30+ large cell lymphoma, non-mycosisfungoides CD30− cutaneous large T-cell lymphoma, pleomorphic T-celllymphoma, Lennert lymphoma, subcutaneous T-cell lymphoma, angiocentriclymphoma, blastic NK-cell lymphoma), prurigo nodularis, lichen planus,contact dermatitis, dyshidrotic eczema, eczema, nummular dermatitis,seborrheic dermatitis, stasis dermatitis, primary localized cutaneousamyloidosis, bullous pemphigoid, skin manifestations of graft versushost disease, pemphigoid, discoid lupus, granuloma annulare, lichensimplex chronicus, pruritus, vulvar/scrotal/perianal pruritus, lichensclerosus, post herpetic neuralgia itch, lichen planopilaris, psoriasis,chronic hand eczema, hidradenitis suppurativa, hypereosinophilicsyndrome, systemic lupus erythematosus, and foliculitis decalvans. Inparticular, atopic dermatitis (Bao et al., JAK-STAT, 2013, 2, e24137),alopecia areata (Xing et al., Nat Med. 2014, 20, 1043-1049) includingsubtypes such as alopecia areata monolocularis, alopecia areatamultilocularis, ophiasis, alopecia areata universalis, alopecia areatatotalis, and alopecia areata barbae, vitiligo (Craiglow et al, JAMADermatol. 2015, 151, 1110-1112), cutaneous T cell lymphoma (Netchiporouket al., Cell Cycle. 2014; 13, 3331-3335), prurigo nodularis (Sonkoly etal., J Allergy Clin Immunol. 2006, 117, 411-417), lichen planus(Welz-Kubiak et al., J Immunol Res. 2015, ID:854747), primary localizedcutaneous amyloidosis (Tanaka et al., Br J Dermatol. 2009, 161,1217-1224), bullous pemphigoid (Feliciani et al., Int J ImmunopatholPharmacol. 1999, 12, 55-61), and dermal manifestations of graft versushost disease (Okiyama et al., J Invest Dermatol. 2014, 134, 992-1000)are characterized by elevation of certain cytokines that signal via JAKactivation. Accordingly, compounds of formula (I) may be able toalleviate associated dermal inflammation or pruritus driven by thesecytokines. In particular, compounds of formula (I), or apharmaceutically acceptable salt thereof, are expected to be useful forthe treatment of atopic dermatitis and other inflammatory skin diseases.

The compounds of formula (I) also possess advantageous solubilityproperties in aqueous and/or organic excipients which facilitateformulation into topical compositions.

The compounds of formula (I) also release an active metabolite, compoundM, thereby increasing overall exposure to JAK inhibitors. Compound Mpossesses favorable properties including as a high clearance andpermeability allowing for rapid systemic clearance and good skinpermeability.

In some embodiments, therefore, the invention provides a method oftreating an inflammatory or autoimmune skin disease in a mammal (e.g., ahuman), comprising applying a pharmaceutical composition comprising acompound of formula (I), or a pharmaceutically acceptable salt thereof,and a pharmaceutical carrier to the skin of the mammal.

In some embodiments, the invention provides a method of treating aninflammatory or autoimmune skin disease in a mammal (e.g., a human),comprising administering a compound of formula (I), or apharmaceutically acceptable salt thereof, to the mammal. In someembodiments, the inflammatory skin disease is atopic dermatitis. In someembodiments, the atopic dermatitis is mild to moderate. In someembodiments, the atopic dermatitis is moderate to severe. In someembodiments, the autoimmune skin disease is alopecia areata.

A compound of formula (I), or a pharmaceutically acceptable saltthereof, may also be used in combination with one or more compounduseful to treat inflammatory skin diseases. In some embodiments, the oneor more compound is a steroid, corticosteroid, antibiotic, Histamine H1receptor antagonist, calcineurin inhibitor, IL-13 antagonist, PDE 4inhibitor, G-protein coupled receptor-44 antagonist, IL-4 antagonist,5-HT 1a receptor antagonist, 5-HT 2b receptor antagonist, Alpha 2adrenoceptor agonist, cannabinoid CB1 receptor antagonist, CCR3chemokine, antagonist, collagenase inhibitor, cytosolic phospholipase A2inhibitor, eotaxin ligand inhibitor, GATA 3 transcription factorinhibitor, Histamine H4 receptor antagonist, IL-10 antagonist, IL-12antagonist, IL-17 antagonist, IL-2 antagonist, IL-23 antagonist, IL-4receptor modulator, IL-15 antagonist, IL-6 antagonist, IL-8 antagonist,IL-9 antagonist, IL-5 antagonist, immunoglobulin E antagonist,immunoglobulin E modulator, interferon gamma receptor antagonist,Interferon gamma ligand, Interleukin 33 ligand inhibitor, Interleukin-31receptor antagonist, Leukotriene antagonist, Liver X receptor agonist,Liver X receptor beta agonist, nuclear factor kappa B inhibitor, OX-40receptor antagonist, PGD2 antagonist, phospholipase A2 inhibitor, SH2domain inositol phosphatase 1 stimulator, thymic stromal lymphoproteinligand inhibitor, TLR modulator, TNF alpha ligand modulator, TLR9 genestimulator, cytotoxic T-lymphocyte protein-4 stimulator, opioid receptorkappa agonist, galectin-3 inhibitor, histone deacetylase-1 inhibitor,histone deacetylase-2 inhibitor, histone deacetylase-3 inhibitor,histone deacetylase-6 inhibitor, histone deacetylase inhibitor,glucocorticoid agonist, Syk tyrosine kinase inhibitor, TrkA receptorantagonist, integrin alpha-4/beta-1 antagonist, Interleukin 1 likereceptor antagonist, Interleukin-1 converting enzyme inhibitor,Interleukin-31 receptor antagonist, KCNA voltage-gated potassiumchannel-3 inhibitor, PDE4B gene inhibitor, Kallikrein 2 inhibitor,sphingosine-1-phosphate receptor-1 agonist, retinal pigment epitheliumprotein stimulator, T cell surface glycoprotein CD28 inhibitor, TGF betaantagonist, vanilloid VR1 antagonist, NK1 receptor antagonist,galectin-3 inhibitor, cytokine receptor antagonist, androgen receptorantagonist, sphingosine 1 phosphate phosphatase 1 stimulator,sphingosine-1-phosphate receptor-1 modulator, sphingosine-1-phosphatereceptor-4 modulator, sphingosine-1-phosphate receptor-5 modulator,Interleukin-1 alpha ligand inhibitor, OX40 ligand inhibitor, Interleukin1 like receptor 2 inhibitor, melanocyte stimulating hormone ligand, CD40ligand receptor antagonist, osteopontin ligand modulator, Interleukin-1beta ligand modulator, I-kappa B kinase beta inhibitor,5-Alpha-reductase inhibitor, 5-Alpha-reductase-1 inhibitor;5-Alpha-reductase-2 inhibitor, sodium channel inhibitor, NACHT LRR PYDdomain protein 3 inhibitor, Wnt ligand modulator, Wnt 7A ligand,melanocortin MC1 receptor agonist, mTOR inhibitor, actin polymerizationmodulator, laminin-5 agonist, metalloprotease-2 modulator,metalloprotease-9 modulator, nuclear factor kappa B inhibitor, thymosinbeta 4 ligand, thymosin receptor agonist, FGF-7 ligand, follistatinagonist, VEGF ligand, MEK-1 protein kinase inhibitor, Ras geneinhibitor, 5-Alpha-reductase inhibitor, alpha 1A adrenoceptorantagonist, Kallikrein 7 inhibitor, cytosolic phospholipase A2inhibitor, elongation factor 2 inhibitor, NAD ADP ribosyltransferasestimulator, Interleukin-2 ligand, nuclear factor kappa B inhibitor,IL-22 antagonist, epidermal growth factor receptor agonist, retinalpigment epithelium protein stimulator, AMP activated protein kinasestimulator, ICE inhibitor, KCNA voltage-gated potassium channel-3inhibitor, G-protein coupled bile acid receptor 1 agonist, or potassiumchannel modulator.

In some embodiments, a compound of formula (I), or a pharmaceuticallyacceptable salt thereof, is administered in combination withbetamethasone, fucidic acid, GR-MD-02, dupilumab, rosiptor acetate,AS-101, ciclosporin, IMD-0354, secukinumab, Actimmune, lebrikizumab,CMP-001, mepolizumab, pegcantratinib, tezepelumab, MM-36, crisaborole,ALX-101, bertilimumab, FB-825, AX-1602, BNZ-1, abatacept, tacrolimus,ANB-020, JTE-052, ZPL-389, ustekinumab, GBR-830, GSK-3772847, ASN-002,remetinostat, apremilast, timapiprant, MOR-106, asivatrep, nemolizumab,fevipiprant, doxycycline, MDPK-67b, desloratadine, tralokinumab,fexofenadine, pimecrolimus, bepotastine, nalfurafine, VTP-38543, Q-301,ligelizumab, RVT-201, DMT-210, KPI-150, AKP-11, E-6005, AMG-0101,AVX-001, PG-102, ZPL-521, MEDI-9314, AM-1030, WOL-071007, MT-0814,betamethasone valerate, SB-011, epinastine, tacrolimus, tranilast,tradipitant, difamilast, LY-3375880, tapinarof, etokimab, clascoterone,etrasimod, bermekimab, KHK-4083, SAR-440340, BI-655130, EDP-1815,EDP-1066, DUR-928, afamelanotide, adriforant, diroleuton, FOL-005,KY-1005, PUR-0110, BTX-1204, ADSTEM, finasteride, BMX-010, BBI-5000,MSB-01, ATI-501, B-244, ASN-008, hypochlorous acid,diphenylcyclopropenone, RG-6149, LY-3454738, SB-414, S1P1 agonist,SM-04554, PL-8177, rapamycin, rose Bengal sodium, tonabacase, omigananpentahydrochloride, desonide, allogeneic mesenchymal stem cell therapy,timbetasin, ASLAN-004, HSC-660, fluocinonide, niclosamide,antroquinonol, dutasteride, tamsulosin, UCA-001, dapsone, brilacidin,BPR-277, anapsos, dutasteride, denileukin diftitox, chanllergen,ARGX-112, PF-06817024, epinastine hydrochloride, IDP-124, nepidermin,roseomonas mucosa-based biotherapy, ENERGI-F701, HAT-1, lotamilast,HY-209, mometasone, melgain, doxycycline, TS-133, icomucret, CRTH2antagonist, ACH-24, fluticasone propionate, CD-4802, minoxidil,finasteride, halometasone, tricomin, or viromed, or any combinationthereof.

In some embodiments, a compound of formula (I), or a pharmaceuticallyacceptable salt thereof, is administered in combination with a steroid,an antibiotic and a moisturizer (Lakhani et al., Pediatric Dermatology,2017, 34, 3, 322-325). In some embodiments, the one or more compound isa gram positive antibiotic, such as mupirocin or fusidic acid.

A compound of formula (I), or a pharmaceutically-acceptable saltthereof, may also be used in combination with gram positive antibiotics,such as mupirocin and fusidic acid, to treat inflammatory skin disease.In one aspect, therefore, the invention provides a method of treating aninflammatory skin disease in a mammal, the method comprising applying acompound of the disclosure, or a pharmaceutically-acceptable saltthereof, and a gram positive antibiotic to the skin of the mammal. Inanother aspect, the invention provides a pharmaceutical compositioncomprising a compound of the disclosure, or apharmaceutically-acceptable salt thereof, a gram positive antibiotic,and a pharmaceutically-acceptable carrier.

In another aspect, therefore, the invention provides a therapeuticcombination for use in the treatment of skin inflammatory disorders, thecombination comprising a compound of formula (I), or a pharmaceuticallyacceptable salt thereof and one or more other therapeutic agents usefulfor treating skin inflammatory disorders. Secondary agent(s), whenincluded, are present in a therapeutically effective amount, i.e. in anyamount that produces a therapeutically beneficial effect whenco-administered with a compound of formula (I), or apharmaceutically-acceptable salt thereof.

Also provided, therefore, is a pharmaceutical composition comprising acompound of formula (I), or a pharmaceutically salt thereof and one ormore other therapeutic agents useful for treating skin inflammatorydisorders.

Further, in a method aspect, the invention provides a method of treatingskin inflammatory disorders, the method comprising administering to themammal a compound of formula (I), or a pharmaceutically acceptable saltthereof, and one or more other therapeutic agents useful for treatingskin inflammatory disorders.

Gastrointestinal Inflammatory Disease

Due to its inhibition of the JAK family of enzymes, compounds of formula(I) are expected to be useful for a variety of gastrointestinalinflammatory indications that include, but are not limited to,ulcerative colitis (proctosigmoiditis, pancolitis, ulcerative proctitisand left-sided colitis), Crohn's disease, collagenous colitis,lymphocytic colitis, Behcet's disease, celiac disease, immune checkpointinhibitor induced colitis, ileitis, eosinophilic esophagitis, graftversus host disease-related colitis, and infectious colitis. Ulcerativecolitis (Reimund et al., J Clin Immunology, 1996, 16, 144-150), Crohn'sdisease (Woywodt et al., Eur J Gastroenterology Hepatology, 1999, 11,267-276), collagenous colitis (Kumawat et al., Mol Immunology, 2013, 55,355-364), lymphocytic colitis (Kumawat et al., 2013), eosinophilicesophagitis (Weinbrand-Goichberg et al., Immunol Res, 2013, 56,249-260), graft versus host disease-related colitis (Coghill et al.,Blood, 2001, 117, 3268-3276), infectious colitis (Stallmach et al., IntJ Colorectal Dis, 2004, 19, 308-315), Behcet's disease (Zhou et al.,Autoimmun Rev, 2012, 11, 699-704), celiac disease (de Nitto et al.,World J Gastroenterol, 2009, 15, 4609-4614), immune checkpoint inhibitorinduced colitis (e.g., CTLA-4 inhibitor-induced colitis; (Yano et al., JTranslation Med, 2014, 12, 191), PD-1- or PD-L1-inhibitor-inducedcolitis), and ileitis (Yamamoto et al., Dig Liver Dis, 2008, 40,253-259) are characterized by elevation of certain pro-inflammatorycytokine levels. As many pro-inflammatory cytokines signal via JAKactivation, compounds described in this application may be able toalleviate the inflammation and provide symptom relief.

In some embodiments, therefore, the disclosure provides a method oftreating a gastrointestinal inflammatory disease in a mammal (e.g., ahuman), comprising administering to the mammal a pharmaceuticalcomposition comprising a pharmaceutically-acceptable carrier and acompound of formula (I) or a pharmaceutically-acceptable salt thereof.

In some embodiments, the disclosure provides a method of treating agastrointestinal inflammatory disease in a mammal (e.g., a human),comprising administering to the mammal a compound of formula (I), or apharmaceutically acceptable salt thereof.

The disclosure further provides a method of treating ulcerative colitisin a mammal, the method comprising administering to the mammal acompound of the disclosure, or a pharmaceutically-acceptable saltthereof, or a pharmaceutical composition comprising apharmaceutically-acceptable carrier and a compound of the disclosure, ora pharmaceutically-acceptable salt thereof.

When used to treat ulcerative colitis, the compound of the inventionwill typically be administered orally in a single daily dose or inmultiple doses per day, although other forms of administration may beused. The amount of active agent administered per dose or the totalamount administered per day will typically be determined by a physician,in the light of the relevant circumstances, including the condition tobe treated, the chosen route of administration, the actual compoundadministered and its relative activity, the age, weight, and response ofthe individual patient, the severity of the patient's symptoms, and thelike.

Suitable doses for treating ulcerative colitis and othergastrointestinal inflammatory disorders are expected to range from about1 to about 400 mg/day of active agent, including from about 5 to about300 mg/day and from about 20 to about 70 mg per day of active agent foran average 70 kg human.

Compounds of formula (I), or a pharmaceutically-acceptable salt thereof,may also be used in combination with one or more agents which act by thesame mechanism or by different mechanisms to effect treatment ofgastrointestinal inflammatory disorders. Useful classes of agents forcombination therapy include, but are not limited to, aminosalicylates,steroids, systemic immunosuppressants, anti-TNFa antibodies, anti-VLA-4antibodies, anti-integrin a437 antibodies, anti-bacterial agents, andanti-diarrheal medicines.

Aminosalicylates that may be used in combination with a compound offormula (I), include, but are not limited to, mesalamine, osalazine andsulfasalazine. Examples of steroids include, but are not limited to,prednisone, prednisolone, hydrocortisone, budesonide, beclomethasone,and fluticasone. Systemic immunosuppressants useful for treatment ofinflammatory disorders include, but are not limited to cyclosporine,azathioprine, methotrexate, 6-mercaptopurine, and tacrolimus. Further,anti-TNFα antibodies, which include, but are not limited to, infliximab,adalimumab, golimumab, and certolizumab, may be used in combinationtherapy. Useful compounds acting by other mechanisms include anti-VLA-4antibodies, such as natalizumab, anti-integrin α₄β₇ antibodies, such asvedolizumab, anti-bacterial agents, such as rifaximin, andanti-diarrheal medicines, such as loperamide. (Mozaffari et al. ExpertOpin. Biol. Ther.2014, 14, 583-600; Danese, Gut, 2012, 61, 918-932; Lamet al., Immunotherapy, 2014, 6, 963-971).

In another aspect, therefore, the disclosure provides a therapeuticcombination for use in the treatment of gastrointestinal inflammatorydisorders, the combination comprising a compound of the disclosure, or apharmaceutically-acceptable salt thereof, and one or more othertherapeutic agents useful for treating gastrointestinal inflammatorydisorders. For example, the disclosure provides a combination comprisinga compound of the disclosure, or a pharmaceutically-acceptable saltthereof, and one or more agents selected from aminosalicylates,steroids, systemic immunosuppressants, anti-TNFα antibodies, anti-VLA-4antibodies, anti-integrin α₄β₇ antibodies, anti-bacterial agents, andanti-diarrheal medicines. Secondary agent(s), when included, are presentin a therapeutically effective amount, i.e. in any amount that producesa therapeutically beneficial effect when co-administered with a compoundof the disclosure, or a pharmaceutically-acceptable salt thereof.

Also provided, therefore, is a pharmaceutical composition comprising acompound of formula (I), or a pharmaceutically-acceptable salt thereof,and one or more other therapeutic agents useful for treatinggastrointestinal inflammatory disorders.

Further, in a method aspect, the disclosure provides a method oftreating gastrointestinal inflammatory disorders, the method comprisingadministering to the mammal a compound of formula (I), or apharmaceutically acceptable salt thereof, and one or more othertherapeutic agents useful for treating gastrointestinal inflammatorydisorders.

Respiratory Diseases

Cytokines which signal through the JAK-STAT pathway, in particular IL-2,IL-3, IL-4, IL-5, IL-6, IL-9, IL-11, IL-13, IL-23, IL-31, IL-27, thymicstromal lymphopoietin (TSLP), interferon-γ (IFNγ) andgranulocyte-macrophage colony-stimulating factor (GM-CSF) have beenimplicated in asthma inflammation and in other inflammatory respiratorydiseases. As described above, compounds of formula (I) have been shownto be a potent inhibitor of Janus kinases and has demonstrated potentinhibition of IL-13 pro-inflammatory cytokines in cellular assays.

The anti-inflammatory activity of JAK inhibitors has been robustlydemonstrated in preclinical models of asthma (Malaviya et al., IntImmunopharmacol, 2010, 10, 829,-836; Matsunaga et al., Biochem andBiophys Res Commun, 2011, 404, 261-267; Kudlacz et al., Eur J Pharmacol,2008, 582, 154-161.) Accordingly, compounds of formula (I), or apharmaceutically acceptable salt thereof, are expected to be useful forthe treatment of inflammatory respiratory disorders such as asthma.Inflammation and fibrosis of the lung is characteristic of otherrespiratory diseases in addition to asthma such as chronic obstructivepulmonary disease (COPD), cystic fibrosis (CF), pneumonitis,interstitial lung diseases (including idiopathic pulmonary fibrosis),acute lung injury, acute respiratory distress syndrome, bronchitis,emphysema, and bronchiolitis obliterans. Compounds of formula (I), or apharmaceutically acceptable salt thereof, therefore, may be useful forthe treatment of chronic obstructive pulmonary disease, cystic fibrosis,pneumonitis, interstitial lung diseases (including idiopathic pulmonaryfibrosis), acute lung injury, acute respiratory distress syndrome,bronchitis, emphysema, bronchiolitis obliterans, chronic lung allograftdysfunction (CLAD), lung transplant rejections, and sarcoidosis.

In one aspect, therefore, the disclosure provides a method of treating arespiratory disease in a mammal (e.g., a human) comprising administeringto the mammal a compound of formula (I), or apharmaceutically-acceptable salt thereof.

In one aspect, the respiratory disease is asthma, chronic obstructivepulmonary disease, cystic fibrosis, pneumonitis, chronic obstructivepulmonary disease (COPD), cystic fibrosis (CF), pneumonitis,interstitial lung diseases (including idiopathic pulmonary fibrosis),acute lung injury, acute respiratory distress syndrome, bronchitis,emphysema, bronchiolitis obliterans, allergic rhinitis or sarcoidosis.In another aspect, the respiratory disease is asthma or chronicobstructive pulmonary disease.

In a further aspect, the respiratory disease is a lung infection, ahelminthic infection, pulmonary arterial hypertension, sarcoidosis,lymphangioleiomyomatosis, bronchiectasis, or an infiltrative pulmonarydisease. In yet another aspect, the respiratory disease is drug-inducedpneumonitis, fungal induced pneumonitis, allergic bronchopulmonaryaspergillosis, hypersensitivity pneumonitis, eosinophilic granulomatosiswith polyangiitis, idiopathic acute eosinophilic pneumonia, idiopathicchronic eosinophilic pneumonia, hypereosinophilic syndrome, Löfflersyndrome, bronchiolitis obliterans organizing pneumonia, orimmune-checkpoint-inhibitor induced pneumonitis.

The disclosure further provides a method of treating a respiratorydisease, the method comprising administering to the mammal apharmaceutical composition comprising a compound of formula (I), or apharmaceutically-acceptable salt thereof and apharmaceutically-acceptable carrier.

Compounds of formula (I), or a pharmaceutically acceptable salt thereof,may also be used in combination with one or more compound useful torespiratory diseases.

Ocular Diseases

Many ocular diseases have been associated with elevations ofproinflammatory cytokines that rely on the JAK-STAT pathway.

Compounds of formula (I), or a pharmaceutically acceptable salt thereof,therefore, may be useful for the treatment of a number of oculardiseases that include, but are not limited to, uveitis, diabeticretinopathy, diabetic macular edema, dry eye disease, age-relatedmacular degeneration, and atopic keratoconjunctivitis.

In particular, uveitis (Horai and Caspi, J Interferon Cytokine Res,2011, 31, 733-744), diabetic retinopathy (Abcouwer, J Clin Cell Immunol,2013, Suppl 1, 1-12), diabetic macular edema (Sohn et al., AmericanJournal of Opthamology, 2011, 152, 686-694), dry eye disease (Stevensonet al, Arch Ophthalmol, 2012, 130, 90-100), retinal vein occlusion(Shchuko et al, Indian Journal of Ophthalmology, 2015, 63(12), 905-911),and age-related macular degeneration (Knickelbein et al, Int OphthalmolClin, 2015, 55(3), 63-78) are characterized by elevation of certainpro-inflammatory cytokines that signal via the JAK-STAT pathway.Accordingly, compounds of formula (I), or a pharmaceutically acceptablesalt thereof, may be able to alleviate the associated ocularinflammation and reverse disease progression or provide symptom relief.

In one aspect, therefore, the disclosure provides a method of treatingan ocular disease in a mammal comprising administering a compound offormula (I), or a pharmaceutically-acceptable salt thereof or apharmaceutical composition comprising a compound of formula (I), or apharmaceutically-acceptable salt thereof and a pharmaceutical carrier tothe eye of the mammal. In one aspect, the ocular disease is uveitis,diabetic retinopathy, diabetic macular edema, dry eye disease,age-related macular degeneration, or atopic keratoconjunctivitis. In oneaspect, the method comprises administering a compound of formula (I), ora pharmaceutically acceptable salt thereof by intravitreal injection.

Compounds of formula (I), or a pharmaceutically acceptable salt thereof,may also be used in combination with one or more compound useful toocular diseases.

Other Diseases

Compounds of formula (I), or a pharmaceutically acceptable salt thereof,may also be useful to treat other diseases such as other inflammatorydiseases, autoimmune diseases or cancers.

Compounds of formula (I), or a pharmaceutically acceptable salt thereof,may be useful to treat oral cavities, oral mucositis and recurrentaphthous stomatitis.

Compounds of formula (I), or a pharmaceutically acceptable salt thereof,may be useful to treat one or more of arthritis, rheumatoid arthritis,juvenile rheumatoid arthritis, transplant rejection, xerophthalmia,psoriatic arthritis, diabetes, insulin dependent diabetes, motor neuronedisease, myelodysplastic syndrome, pain, sarcopenia, cachexia, septicshock, systemic lupus erythematosus, leukemia, chronic lymphocyticleukemia, chronic myelocytic leukemia, acute lymphoblastic leukemia,acute myelogenous leukemia, ankylosing spondylitis, myelofibrosis,B-cell lymphoma, hepatocellular carcinoma, Hodgkins disease, breastcancer, Multiple myeloma, melanoma, squamous cell carcinoma, non-Hodgkinlymphoma, non-small-cell lung cancer, ovarian clear cell carcinoma,ovary tumor, pancreas tumor, polycythemia vera, Sjoegrens syndrome, softtissue sarcoma, sarcoma, splenomegaly, T-cell lymphoma, and thalassemiamajor.

The disclosure, thereof, provides a method of treating these diseases ina mammal comprising administering a compound of formula (I), or apharmaceutically-acceptable salt thereof or a pharmaceutical compositioncomprising a compound of formula (I), or a pharmaceutically-acceptablesalt thereof and a pharmaceutical carrier to the mammal.

In the previous paragraphs, when used in combination therapy, the agentsmay be formulated in a single pharmaceutical composition, as disclosedabove, or the agents may be provided in separate compositions that areadministered simultaneously or at separate times, by the same or bydifferent routes of administration. When administered separately, theagents are administered sufficiently close in time so as to provide adesired therapeutic effect. Such compositions can be packaged separatelyor may be packaged together as a kit. The two or more therapeutic agentsin the kit may be administered by the same route of administration or bydifferent routes of administration.

EXAMPLES

The following synthetic and biological examples are offered toillustrate the invention, and are not to be construed in any way aslimiting the scope of the invention. In the examples below, thefollowing abbreviations have the following meanings unless otherwiseindicated. Abbreviations not defined below have their generally acceptedmeanings.

ACN=acetonitrile

Bn=benzyl

Boc=tert-Butyloxycarbonyl

d=day(s)

DIPEA=N,N-diisopropylethylamine

DMF=N,N-dimethylformamide

DMSO=dimethyl sulfoxide

EtOAc=ethyl acetate

EtOH=ethyl alcohol

h=hour(s)

HATU=N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uroniumhexafluorophosphate

IPA=isopropyl alcohol

MeOH=methanol

min=minute(s)

NMP=N-methylpyrrolidone

RT=room temperature

TEA=triethylamine

THF=tetrahydrofuran

TFA=trifluoroacetic acid

Reagents and solvents were purchased from commercial suppliers (Aldrich,Fluka, Sigma, etc.), and used without further purification. Progress ofreaction mixtures was monitored by thin layer chromatography (TLC),analytical high performance liquid chromatography (anal. HPLC), and/ormass spectrometry. Reaction mixtures were worked up as describedspecifically in each reaction; commonly they were purified by extractionand other purification methods such as temperature-, andsolvent-dependent crystallization, and precipitation. In addition,reaction mixtures were routinely purified by column chromatography or bypreparative HPLC, typically using C18 or BDS column packings andconventional eluents. Typical preparative HPLC conditions are describedbelow.

Characterization of reaction products was routinely carried out by massand ¹H-NMR spectrometry. For NMR analysis, samples were dissolved indeuterated solvent (such as CD₃OD, CDCl₃, or d₆-DMSO), and ¹H-NMRspectra were acquired with a Varian Gemini 2000 instrument (400 MHz)under standard observation conditions. Mass spectrometric identificationof compounds was performed by an electrospray ionization method (ESMS)with an Applied Biosystems (Foster City, Calif.) model API 150 EXinstrument or a Waters (Milford, Mass.) 3100 instrument, coupled toautopurification systems.

Unless otherwise indicated the following conditions were used forpreparative HPLC purifications.

Column: C18, 5 μm 21.2×150 mm or C18, 5 μm 21×250 mm or C14, 5 μm 21×150mm

Column temperature: Room Temperature

Flow rate: 20.0 mL/min

Mobile Phases: A=Water+0.05% TFA

B=ACN+0.05% TFA,

Injection volume: (100-1500 μL)

Detector wavelength: 214 nm

Crude compounds were dissolved in 1:1 water:acetic acid at about 50mg/mL. A 4 minute analytical scale test run was carried out using a2.1×50 mm C18 column followed by a 15 or 20 minute preparative scale runusing 100 μL injection with the gradient based on the % B retention ofthe analytical scale test run. Exact gradients were sample dependent.Samples with close running impurities were checked with a 21×250 mm C18column and/or a 21×150 mm C14 column for best separation. Fractionscontaining desired product were identified by mass spectrometricanalysis.

Analytic HPLC Conditions

Method A

Column: LUNA C18 (2), 150×4.60 mm, 3 μm

Column temperature: 37° C.

Flow rate: 1.0 mL/min

Injection volume: 5 μL

Sample preparation: Dissolve in 1:1 ACN:water

Mobile Phases: A=Water:ACN:TFA (98:2:0.05)

B=Water:ACN:TFA (2:98:0.05)

Detector wavelength: 250 nm

Gradient: 32 min total (time (min)/% B): 0/2, 10/20, 24/90, 29/90, 30/2,32/2

Method B

Column: LUNA C18 (2), 150×4.60 mm, 3 μm

Column temperature: 37° C.

Flow rate: 1.0 mL/min

Injection volume: 10 μL

Sample preparation: Dissolve in 1:1 ACN:water

Mobile Phases: A=Water:ACN:TFA (98:2:0.05)

B=Water:ACN:TFA (10:90:0.05)

Detector wavelength: 254 nm

Gradient: 35 min total (time (min)/% B): 0/2, 20/25, 23/90, 26/90, 27/2,35/2

Method C

Column: Poroshell 120 SB-Aq, 150 mm by 4.6 mm, 2.7 micron part#683975-914

Column temperature: 35° C.

Flow rate: 1.0 mL/min

Injection volume: 5 μL

Sample preparation: Dissolve in 50:MPB:50MPA

Mobile Phases: A=Acetonitrile:Water:Trifluoroacetic acid (1:99:0.20)

B=Acetonitrile:Water:Trifluoroacetic acid (90:10:0.20)

Gradient:

Time, min % A % B 0.0 98.0 2.0 16.0 40.0 60.0 22.0 0.0 100.0 25.0 0.0100.0 25.1 98.0 2.0 30.0 98.0 2.0

Preparation 1: tert-butyl((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)carbamate

Step 1: Five reactions were carried out in parallel. To a solution ofcompound 1-1 (2.00 kg, 13.7 mol, 1.00 eq) in dioxane (5.00 L) and water(20.0 L) was added glutaraldehyde (2.06 kg, 20.5 mol, 1.5 eq) andphenylmethanamine (1.54 kg, 14.4 mol, 1.05 eq) drop-wise at 10° C. Afteraddition, the reaction mixture was stirred at 20° C. for 16 h. TLC(petroleum ether:ethyl acetate=5:1, product R_(f)=0.40) and LCMSindicated the reaction was complete. The pH value of the reactionmixture was adjusted to 2 with concentrated HCl (12 N) at 20° C. Afteraddition, the reaction mixture was heated to 60° C. and stirred for 1 h.After cooling to 10° C., ethyl acetate (10.0 L) was added to themixture. Then the pH value of the mixture was adjusted to 10 by addingan aqueous solution of sodium hydroxide (12 N) at 10° C. The mixture wasstirred for 10 min. The organic layer was separated. The aqueous layerwas extracted with ethyl acetate (3.00 L). The combined organic layerswere washed with brine (4.00 L), dried over sodium sulfate, andfiltered. The organic layer for the five parallel reactions was combinedand concentrated. The residue was purified by column chromatography(SiO₂, petroleum ether:ethyl acetate=30:1-2:1) to give compound 1-2(10.0 kg, 51.5% yield, 97% purity). (m/z): [M+H]⁺ calcd for C₁₅H₁₉NO230.15 found 230.0. ¹H NMR: 400 MHz DMSO-d₆ δ7.24-7.41 (m, 5H), 3.88 (s,2H), 3.20-3.21 (m, 2H), 2.73-2.79 (m, 2H), 2.07 (d, J=16.4 Hz, 2H),1.75-1.84 (m, 2H), 1.45-1.50 (m, 3H), 1.24-1.36 (m, 1H).

Step 2: Three reactions were carried out in parallel. To a solution ofcompound 1-2 (3.00 kg, 13.1 mol, 1.0 eq) in ethyl acetate (24.0 L) andwater (9.00 L) was added CH₃COOK (2.05 kg, 20.9 mol, 1.6 eq) andNH₂OH—HCl (1.82 kg, 26.2 mol, 2.0 eq) at 20° C. The suspension washeated to 45° C. and stirred for 16 h. TLC (petroleum ether:ethylacetate=2:1, product R_(f)=0.30) and LCMS indicated the reaction wascomplete. The pH value of the suspension was adjusted to 8 withsaturated sodium bicarbonate solution, then diluted with water (15.0 L)and ethyl acetate (10.0 L). The organic layer was separated. The aqueouslayer was extracted with ethyl acetate (10.0 L×3). The organic layer ofthe three reactions was combined, dried over sodium sulfate, filteredand concentrated. The crude product was diluted with n-heptane (12.0 L),and stirred for 12 h. The solid was collected by filtration to givecompound 1-3 (8.00 kg, 83.4% yield). (m/z): [M+H]⁺ calcd for C₁₅H₂₀N₂O245.16 found 245.1. ¹H NMR: 400 MHz DMSO-d₆ 10.16 (s, 1H), 7.22-7.38 (m,5H), 3.83 (s, 2H), 2.97 (br s, 2H), 2.87 (d, J=16.0 Hz, 1H), 2.60-2.62(m, 1H), 2.20-2.25 (m, 1H), 2.09-2.13 (m, 1H), 1.72-1.85 (m, 3H),1.39-1.49 (m, 3H).

Step 4: Forty-five reactions were carried out in parallel. To a solutionof compound 1-3 (160 g, 655 mmol, 1.0 eq) in n-PrOH (3.20 L) at 110° C.was added Na (181 g, 7.86 mol, 12 eq) in portions over 3 h. The mixturewas stirred at 110° C. for 2 h. TLC (petroleum ether:ethyl acetate=2:1,SM R_(f)=0.40) indicated the reaction was complete. The mixture wascooled to 70° C., poured into ice water (4.00 L). The aqueous layer wasextracted with ethyl acetate (1.00 L×2). The combined organic layer ofthe forty-five reactions was washed with brine (20.0 L), dried oversodium sulfate, filtered and concentrated. The residue was diluted withn-hexane (12.0 L), stirred for 12 h. The suspension was filtered to getfiltrate. The filtrate was concentrated to give compound 1-4 (6.00 kg,88.4% yield) as a yellow oil. ¹H NMR 400 MHz DMSO-d₆: δ7.18-7.35 (m,5H), 3.76 (s, 2H), 3.26-3.35 (m, 1H), 2.76 (s, 2H), 1.86-1.90 (m, 2H),1.67-1.73 (m, 2H), 1.54-1.59 (m, 5H), 1.41-1.45 (m, 3H).

Step 5: Two reactions were carried out in parallel. To a solution ofcompound 1-4 (2.10 kg, 9.12 mol, 1.1 eq) in dioxane (12.6 L) and water(1.26 L) was added Et3N (1.01 kg, 10.0 mol, 1.1 eq) and (Boc)₂O (2.19kg, 10.0 mol, 1.1 eq) drop-wise at 0° C., with the temperature below 20°C. The mixture was heated to 40° C. and stirred for 10 h. TLC (petroleumether:ethyl acetate=2:1, product R_(f)=0.40) showed the reaction wascomplete. The mixture was cooled to 10° C., filtered to get filter cake.The filtrate was concentrated. The filter cake was washed with n-hexane(3.00 L) to give compound 1-5 (4.00 kg, 66.4% yield) as a white solid.¹H NMR: 400 MHz DMSO-d₆: δ7.28-7.33 (m, 4H), 7.19-7.22 (m, 1H), 6.64 (d,J=8.0 Hz, 1H), 4.10-4.17 (m, 1H), 3.77 (s, 2H), 2.77 (s, 2H), 1.88-1.90(m, 2H), 1.72-1.75 (m, 3H), 1.57-1.61 (m, 3H), 1.43-1.48 (m, 2H), 1.38(s, 9H).

Step 6: Four reactions were carried out in parallel. To a suspension ofcompound 1-5 (1.50 kg, 4.54 mol, 1.0 eq) in DMF (13.5 L) was added NaH(272 g, 6.81 mol, 60% purity, 1.5 eq) portion-wise at 0° C. under N₂.The suspension was naturally warmed to 25° C. and stirred for 30 min.After it was cooled down to 0° C., MeI (773 g, 5.45 mol, 1.2 eq) wasadded drop-wise to the suspension. The reaction mixture was naturallywarmed to 25° C. and stirred for 12 h. TLC (petroleum ether:ethylacetate=5:1, product R_(f)=0.50) and LCMS showed the reaction wascomplete. The mixture was poured into ice water (30.0 L), extracted withethyl acetate (9.00 L, 3.00 L). The combined organic layer of the fourreactions was washed with ice water (20.0 L), brine (10.0 L), dried oversodium sulfate, filtered and concentrated to give compound 1-6 (6.00 kg,crude) as a yellow oil. The crude product was used for the next step. ¹HNMR: 400 MHz DMSO-d₆ δ7.21-7.37 (m, 5H), 4.87 (br s, 1H), 3.80 (s, 2H),2.86 (s, 2H), 2.68 (s, 3H), 1.64-1.99 (m, 6H), 1.40-1.49 (m, 13H).(m/z): [M+H]⁺ calcd for C₂₁H₃₂N₂O₂ 344.25 found 345.2.

Step 7: Thirty-nine reactions were carried out in parallel. To asolution of compound 1-6 (150 g, 435 mmol, 1.0 eq) in IPA (500 mL) andTHF (500 mL) was added Pd(OH)₂/C (70 g, 40% purity). The suspension wasdegassed under vacuum and purged with Hz several times. The mixture wasstirred under Hz (50 psi) at 25° C. for 16 h. TLC (petroleum ether:ethylacetate=5:1, SM R_(f)=0.50) and LCMS indicated the reaction wascomplete. The thirty-nine reactions were combined. The mixture wasfiltered to get filtrate. The filter cake was washed with IPA/THF (1:1,25.0 L). The combined filtrate was concentrated to give compound 1-7(3.85 kg, crude) as a light yellow oil. The crude product was used forthe next step directly. (m/z): [M+H]⁺calcd for C₁₄H₂₆N₂O₂ 255.20 found255.1. ¹H NMR: 400 MHz DMSO-d₆ δ4.88 (br s, 1H), 3.08 (s, 2H), 2.60 (s,3H), 1.73-1.76 (m, 5H), 1.51-1.61 (m, 5H), 1.39 (s, 9H).

Step 8: Four reactions were carried out in parallel. To a solution ofcompound 1-7 (750 g, 2.95 mol, 1.0 eq) in 2-methyl tetrahydrofuran (3.00L) was added pyridine (466 g, 5.90 mol, 2.0 eq) and ethanesulfonylchloride (398 g, 3.10 mol, 1.05 eq) drop-wise at 0° C. under N₂. Themixture was warmed to 25° C. and stirred for 3 h. TLC (petroleumether:ethyl acetate=2:1, product R_(f)=0.50) indicated the reaction wascomplete. The four reactions were combined. The mixture was quenchedwith ice water (10.0 L). The organic layer was separated, washed with0.5 N HCl (3.00 L×2). The combined aqueous layer was extracted withethyl acetate (3.00 L), the organic layer was washed with 0.5 N HCl (500mL) again. The combined organic layer was washed with brine (5.00 L),dried over sodium sulfate, filtered and concentrated to give compound1-8 (2.20 kg, crude) as a yellow oil. The crude product was used in thenext step. ¹H NMR: 400 MHz DMSO-d₆ δ4.94 (br s, 1H), 3.98 (s, 2H), 3.10(q, J=7.2 Hz, 2H), 2.58 (s, 3H), 1.83-1.91 (m, 5H), 1.56-1.71 (m, 5H),1.40 (s, 9H), 1.19 (t, J=7.2 Hz, 3H).

Step 9: Four reactions were carried out in parallel. To a solution ofcompound 1-8 (550 g, 1.59 mol, 1.0 eq) in EtOAc (2.75 L) was addedHC1/EtOAc (4 M, 3.0 eq) drop-wise at 25° C. The mixture was stirred at25° C. for 12 h. TLC (petroleum ether: ethyl acetate=2:1, SM R_(f)=0.50)showed the reaction was complete. The four reactions were combined. Themixture was filtered to get filter cake to give compound 1-9 (1.25 kg,crude, HCl) as a yellow solid. ¹H NMR: 400 MHz DMSO-d₆ δ9.04 (s, 1H),4.02 (s, 2H), 3.88-3.94 (m, 1H), 3.09 (q, J=7.2 Hz, 2H), 2.09-2.14 (m,2H), 1.61-1.84 (m, 8H), 1.19 (t, J=7.2 Hz, 3H).

Preparation 2:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanolM

Step 1: A solution of compound 2-1 (1.00 kg, 5.74 mol, 1.0 eq) inethanol (15.0 L) with saturated HCl (1.40 kg, 38.4 mol) was stirred at90° C. for 60 h. HPLC showed one main peak was detected. The reactionmixture was filtered. The filter cake was collected to give compound 2-2(1.00 kg, 81.8% yield, 98.8% purity) as a white solid. ¹H NMR: 400 MHzDMSO-d₆ δ11.82 (br s, 1H), 10.82 (br s, 1H), 4.31 (q, J=7.2 Hz, 2H),1.27 (t, J=6.8 Hz, 3H).

Step 2: Five reactions were carried out in parallel. To a solution ofcompound 2-2 (560 g, 2.77 mol, 1.0 eq) in POCl₃ (1.68 L) was added N,N-diethylaniline (289 g, 1.94 mol, 0.7 eq). The mixture was stirred at140° C. for 12 h. TLC (petroleum ether:ethyl acetate=10:1, productR_(f)=0.50) indicated compound 2-2 was consumed completely. The fivereactions were combined. The reaction mixture was concentrated underreduced pressure to give a residue. The residue was diluted with ethylacetate (25.0 L). The solution was poured into crushed ice (25.0 L). Thewater phase was extracted with ethyl acetate (25.0 L). The combinedorganic layers were washed with saturated sodium carbonate solution(10.0 L×2), dried over sodium sulfate, filtered and concentrated underreduced pressure to give a residue. The residue was purified by columnchromatography (SiO₂, petroleum ether:ethyl acetate=1:0-50:1) to givecompound 2-3 (2.00 kg) as a brown liquid. ¹H NMR: 400 MHz CDCl₃ 4.51 (q,J=7.2 Hz, 2H), 1.44 (t, J=7.2 Hz, 3H).

Step 3: Four reactions were carried out in parallel. A mixture ofcompound 2-3 (480 g, 2.01 mol, 1.0 eq), compound 2-4 (224 g, 2.31 mol,1.15 eq), DIPEA (519 g, 4.02 mol, 2.0 eq) in ethanol (2.60 L) wasdegassed and purged with N₂ for 3 times, and then the mixture wasstirred at 25° C. for 4 h under N₂ atmosphere. TLC (petroleum ether:ethyl acetate=10:1) indicated compound 2-3 was consumed completely. TLC(petroleum ether:ethyl acetate=1:1, product R_(f)=0.40) indicated onenew spot formed. The four reactions were combined. The reaction mixturewas filtered and the filter cake was collected. The filtrate wasconcentrated under reduced pressure to give a residue. The residue wastriturated with water (38.0 L) and filtered. The filter cake (300 g) wastriturated with ethanol (600 mL) and filtered. The two filter cakes werecombined to give compound 2-5 (1.50 kg, 62.2% yield) as a yellow solid.¹H NMR: 400 MHz DMSO-d₆ δ12.31 (s, 1H), 10.76 (s, 1H), 6.38 (s, 1H),4.35 (q, J=7.2 Hz, 2H), 2.27 (s, 3H), 1.30 (t, J=7.2 Hz, 3H).

Step 4: Four reactions were carried out in parallel. A solution ofcompound 2-5 (254 g, 848 mmol, 1.0 eq), compound 1-9 (300 g, 1.06 mol,HCl, 1.25 eq) and DIPEA (548 g, 4.24 mol, 5.0 eq) in DMSO (600 mL) wasstirred at 130° C. for 16 h. TLC (ethyl acetate:petroleum ether=2:1,R_(f)=0.30) and LCMS showed ˜9% of the starting material remained. Themixture was cooled to 25° C. The four reactions were combined, pouredinto ice water (12.0 L). A yellow precipitate was formed. The solid wascollected by filtration to give compound 2-6 (1.50 kg, ˜76% purity) as ayellow solid. (m/z): [M+H]⁺ calcd for C₂₂H₃₂FN₇O₄S 510.22 found 510.2.

A suspension of compound 2-6 (440 g, 656 mmol, ˜76% purity) in ethanol(1.10 L) was heated to 95° C. until the solid was dissolved. Thesolution was cooled to 25° C. and stirred for 12 h. HPLC showed ˜96.9%purity. The three reactions were combined. The suspension was filteredto get the filter cake to give compound 2-6 (˜570 g, 96.9% purity) as alight yellow solid. The product was used for the next step directly. ¹HNMR: 400 MHz DMSO-d₆ δ12.12 (s, 1H), 9.73 (s, 1H), 6.35 (s, 1H), 5.59(br s, 1H), 4.32 (m, 2H), 4.02 (s, 2H), 3.13 (q, J=7.2 Hz, 2H), 2.83 (s,3H), 2.20 (s, 3H), 1.94 (s, 3H), 1.64-1.73 (m, 5H), 1.76-1.87 (m, 5H),1.29 (t, J=7.2 Hz, 3H), 1.21 (t, J=7.2 Hz, 3H).

Step 5: Five reactions were carried out in parallel. To a solution ofcompound 2-6 (130 g, 255 mmol, 1.0 eq) in tetrohydrofuran (3.25 L) andethanol (3.25 L) was added NaBH₄ (77.2 g, 2.04 mol, 8.0 eq) and CaCl₂(113 g, 1.02 mol, 4.0 eq) portion-wise at 0° C. The mixture was warmedto 10° C. and stirred for 2 h. TLC (ethyl acetate:petroleum ether=3:1,product R_(f)=0.20) showed the reaction was complete. The five reactionswere combined. The mixture was quenched by saturated sodium carbonatesolution (6.00 L), diluted with ethyl acetate (15.0 L) and stirred for0.5 h. The suspension was filtered to get filtrate. The organic layerwas separated, and aqueous layer was extracted with ethyl acetate (5.00L×2). The combined organic layer was washed with brine (5.00 L), driedover sodium sulfate, filtered and concentrated to give compound M (500g, crude) as a light yellow solid.

Purification: Five reactions were carried out in parallel. A suspensionof M (100 g, 210 mmol) in ethanol (3.00 L) was heated to 95° C. untilthe solid was dissolved. The solution was cooled to 25° C. and stirredfor 12 h, a lot of precipitate formed. HPLC showed 100% purity. The fivereactions were combined. The solid was collected by filtration to give atotal of 330 g of compound M (99.3% purity) as a light yellow solid(crystalline Form I). (m/z): [M+H]⁺ calcd for C₂₀H₃₀FN₇O₃S 468.21 found468.3. ¹H NMR: 400 MHz DMSO-d₆ δ12.02 (s, 1H), 9.29 (s, 1H), 6.34 (s,1H), 5.61 (br s, 1H), 5.02 (t, J=6.8 Hz, 1H), 4.33 (d, J=4.0 Hz, 2H),4.02 (s, 2H), 3.12 (q, J=7.2 Hz, 2H), 2.84 (s, 3H), 2.19 (s, 3H),1.82-2.01 (m, 3H), 1.63-1.74 (m, 5H), 1.21 (t, J=7.2 Hz, 3H).

Preparation 3: ethyl 5-fluoro-2,6-dihydroxypyrimidine-4-carboxylate

A solution of 5-fluoro-2,6-dihydroxypyrimidine-4-carboxylic acid (20.4g, 120 mmol) in DMF (200 mL) was treated with DBU (18.7 g, 123 mmol) andwas stirred for 0.5 h at 25° C. Then EtI (19.2 g, 123 mmol) was addedand the resulting solution was heated to 60° C. for 3 hours. H₂O (1000mL) was added to the mixture, and the resulting precipitate wascollected by filtration, washed with H₂O (200 mL), and dried to giveethyl 5-fluoro-2,6-dihydroxypyrimidine-4-carboxylate (19 g, 80% yield).

Preparation 4: ethyl 2,6-dichloro-5-fluoropyrimidine-4-carboxylate

A mixture of ethyl 5-fluoro-2,6-dihydroxypyrimidine-4-carboxylate (5 g,24.8 mmol), PhNEt₂ (2.58 g, 17.3 mmol), POCl₃ (130 g, 855.9 mmol) washeated to 100° C. for 4 hours. Then the reaction mixture was cooled toroom temperature and poured into ice water (500 mL). The aqueous layerwas extracted with EtOAc (1000 mL) and the organic layer was washed withsat. NaHCO₃ (200 mL), brine (200 mL), dried over Na₂SO₄, filtered, andconcentrated under vacuum. The residue was purified by columnchromatography (80 g column; 0-50% EtOAc in hexanes) to give ethyl2,6-dichloro-5-fluoropyrimidine-4-carboxylate as yellow oil (3.8 g,65%).

Preparation 5: ethyl2-chloro-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxylate

A mixture of ethyl 2,6-dichloro-5-fluoropyrimidine-4-carboxylate (3.8 g,16 mmol), 5-methyl-1H-pyrazol-3-amine (1.86 g, 19 mmol), and DIPEA (4 g,32 mmol) in EtOH (100 mL) was stirred at r.t. for 2 h. The reactionmixture was concentrated under vacuum. Then water (500 mL) was added andthe reaction mixture was filtered and the filter cake was washed with100 mL of H₂O, and dried in vacuo to give ethyl2-chloro-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxylate(3.8 g 80% yield).

Preparation 6: tert-butyl(1R,3s,5S)-3-((4-(ethoxycarbonyl)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate

A mixture of ethyl2-chloro-5-fluoro-6-((5-methyl-1H-pyrazol-3-ylamino)pyrimidine-4-carboxylate(1.7 g, 5.684 mmol), tert-butyl(1R,3s,5S)-3-(methylamino)-9-azabicyclo[3.3.1]nonane-9-carboxylate (2.17g, 8.527 mmol), and DIPEA (1.47 g, 11.368 mmol) in DMSO (50 mL) washeated to 110° C. for 18 h. The reaction mixture was poured into water(200 mL) and the reaction mixture was filtered and the filter cake waswashed with 200 mL of H₂O and dried in vacuum to give crude tert-butyl(1R,3s,5S)-3-((4-(ethoxycarbonyl)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate(3.5 g, crude). (m/z): [M+H]⁺ calcd for C₂₅H₃₇FN₇O₄ 518.29 found 518.2.

Preparation 7: tert-butyl(1R,3s,5S)-3-((5-fluoro-4-(hydroxymethyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate

A mixture oftert-butyl(1R,3s,5S)-3-((4-(ethoxycarbonyl)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate(3.5 g, 7 mmol), NaBH₄ (2.1 g, 56 mmol), and CaCl₂ (3.1 g, 28 mmol) in amixture of EtOH (50 mL) and THF (50 mL) was stirred overnight at 25° C.The reaction mixture was quench with Na₂CO₃ (aq) (80 mL) and H₂O (80mL), the aqueous layer was extracted with EtOAc (100 mL×3) and thecombined organic layers were washed with brine, dried over Na₂SO₄, andconcentrated under vacuum. The residue was purified by prep-HPLC to givetert-butyl(1R,3s,5S)-3-((5-fluoro-4-(hydroxymethyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate(1.4 g, 44%). (m/z): [M+H]⁺ calcd for C₂₃H₃₅FN₇O₃ 476.28 found 476.3.

Preparation 8:(2-(((1R,3s,5S)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol

A solution oftert-butyl(1R,3s,5S)-3-((5-fluoro-4-(hydroxymethyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate(1.4 g, 2.95 mmol) in HCl/dioxane (50 mL) was stirred at 25° C. for 4 h.The reaction mixture was filtered and the filter cake was washed with100 mL of EtOAc and dried in vacuum to give(2-(((1R,3s,5S)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(1.4 g, 100%). (m/z): [M+H]⁺ calcd for C₁₈H₂₇FN₇O 376.23 found 376.2.

Preparation 9:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol

(2-((1R,3s,5S)-9-azabicyclo[3.3.1]nonan-3-yl(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(95 mg, 0.253 mmol) was dissolved in Pyridine (4.0 ml) and treated withethanesulfonyl chloride (0.024 ml, 0.253 mmol). The reaction mixture wasstirred for 2 hours and subsequently concentrated in vacuo. The cruderesidue was dissolved in 3 mL of a 1:1 mixture of acetic acid/water,filtered to remove particulate, and purified by preparative HPLC(Agilent Dynamax 250×21.4 mm 10 μm, 15 mL/min, 2-50% ACN+0.05% TFA/ACN)using a 2-50% gradient of ACN in water with 0.05% TFA). Pure fractionswere combined and lyophilized to provide the TFA salt of the titlecompound (12.92 mg, 8.8% yield, 99.9% purity). (m/z): [M+H]⁺ calcd forC₂₀H₃₁FN₇O₃S 468.22 found 468.

General Procedure 1: Acylation of Primary Alcohol

An acyl chloride (3.0 equiv., 0.77 mmol) was added to a mixture of(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(1.0 equiv., 0.12 g, 0.26 mmol), diisopropylethylamine (3 equiv., 0.134mL, 0.77 mmol), and N,N-dimethylaminopyridine (0.1 equiv., 3.1 mg, 0.026mmol) in DMF (3.0 mL). The mixture was stirred at ambient temperaturefor 1.5 h. Hydrazine (4.5 equiv., 0.036 mL, 1.2 mmol) was added, and themixture was stirred at ambient temperature for 30 min. Acetic acid (10equiv., 0.147 mL, 2.6 mmol) was added, and the mixture was concentrated.

General Procedure 2: Formation of Free-Base Amorphous Solids

The trifluoroacetic acid salt of the desired compound was dissolved inMeOH (2.5 mL), and immobilized bicarbonate resin (PL-HCO₃ MP Resin (100Å, 2.08 mmol/g, 150-300 μm)) was added. The mixture was agitated on amechanical shaker at ambient temperature for 2 h. The resin was removedby filtration, and the filtrate was concentrated to give free-baseamorphous solid.

Example 1: Preparation of(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methylacetate

Compound 1 was prepared following general procedure 1, using acetylchloride (0.055 mL, 0.77 mmol). The remaining residue was purified bynormal-phase column chromatography (silica gel flash column, 0-10%MeOH/DCM), followed by reverse-phase column chromatography (RediSep PrepC18 column, 20-60% H₂O/CH₃CN). The free-base form of the desiredcompound was generated following general procedure 2, using immobilizedbicarbonate resin (0.38 g, 0.78 mmol) to afford the desired product asan amorphous white solid (0.062 g, 0.12 mmol, 47% yield). LC-MS: m/z[M+H]⁺=510.2 (calculated: 510.22); ¹H NMR: 400 MHz DMSO-d₆ δ12.04 (s,1H), 9.44 (s, 1H), 6.32 (s, 1H), 5.58 (s, 1H), 4.97 (s, 2H), 4.02 (app.s, 2H), 3.12 (q, J=7.3 Hz, 2H), 2.81 (s, 3H), 2.19 (s, 3H), 2.08 (s,3H), 1.96 (m, 3H), 1.86 (m, 2H), 1.67 (m, 5H), 1.21 (t, J=7.3 Hz, 3H).

Example 2: Preparation of(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methylpropionate

Compound 2 was prepared following general procedure 1, using propionylchloride (0.067 mL, 0.77 mmol). The remaining residue was purified byreverse-phase column chromatography (RediSep Prep C18 column, 25-60%H₂O/CH₃CN). The free-base form of the desired compound was generatedfollowing general procedure 2, using immobilized bicarbonate resin (0.45g, 0.94 mmol) to afford the desired product as an amorphous white solid(0.087 g, 0.16 mmol, 62% yield). LC-MS: m/z [M+H]⁺=524.1 (calculated:524.24); ¹H NMR: 400 MHz DMSO-d₆ δ12.04 (s, 1H), 9.42 (s, 1H), 6.33 (s,1H), 5.58 (s, 1H), 4.99 (s, 2H), 4.02 (app. s, 2H), 3.12 (q, J=7.3 Hz,2H), 2.80 (s, 3H), 2.38 (q, J=7.5 Hz, 2H), 2.19 (s, 3H), 1.96 (m, 3H),1.85 (m, 2H), 1.67 (m, 5H), 1.21 (t, J=7.3 Hz, 3H), 1.06 (t, J=7.5 Hz,3H).

Example 3:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methylbutyrate

Compound 3 was prepared following general procedure 1, using butyrylchloride (0.081 mL, 0.77 mmol). The remaining residue was purified bynormal-phase column chromatography (silica gel flash column, 0-10%MeOH/DCM), followed by reverse-phase column chromatography (RediSep PrepC18 column, 30-70% H₂/CH₃CN). The free-base form of the desired compoundwas generated following general procedure 2, using immobilizedbicarbonate resin (0.317 g, 0.66 mmol) to afford the desired product asan amorphous white solid (0.058 g, 0.11 mmol, 42% yield). LC-MS: m/z[M+H]⁺=538.2 (calculated: 538.25); ¹H NMR: 400 MHz DMSO-d₆δ12.05 (s,1H), 9.46 (s, 1H), 6.33 (s, 1H), 5.58 (s, 1H), 4.99 (s, 2H), 4.01 (app.s, 2H), 3.12 (q, J=7.2 Hz, 2H), 2.80 (s, 3H), 2.34 (t, J=7.3 Hz, 2H),2.19 (s, 3H), 1.96 (m, 3H), 1.85 (m, 2H), 1.66 (m, 5H), 1.56 (dt, J=14.6Hz, 7.3 Hz, 2H), 1.21 (t, J=7.2 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H).

Example 4:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methylpentanoate

Compound 4 was prepared following general procedure 1, using valeroylchloride (0.093 mL, 0.77 mmol). The remaining residue was purified bynormal-phase column chromatography (silica gel flash column, 0-10%MeOH/DCM), followed by reverse-phase column chromatography (RediSep PrepC18 column, 40-75% H₂/CH₃CN). The free-base form of the desired compoundwas generated following general procedure 2, using immobilizedbicarbonate resin (0.225 g, 0.47 mmol) to afford the desired product asan amorphous white solid (0.045 g, 0.082 mmol, 32% yield). LC-MS: m/z[M+H]⁺=552.3 (calculated: 552.27); ¹H NMR: 400 MHz DMSO-d₆ δ12.04 (s,1H), 9.42 (s, 1H), 6.33 (s, 1H), 5.59 (s, 1H), 4.98 (s, 2H), 4.02 (app.s, 2H), 3.12 (q, J=7.1 Hz, 2H), 2.80 (s, 3H), 2.36 (t, J=7.4 Hz, 2H),2.19 (s, 3H), 1.96 (m, 3H), 1.86 (m, 2H), 1.66 (m, 5H), 1.53 (m, 2H),1.31 (dt, J=14.9 Hz, 7.3 Hz, 2H), 1.20 (t, J=7.2 Hz, 3H), 0.86 (t, J=7.3Hz, 3H).

Example 5:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methylheptanoate

Compound 5 was prepared following general procedure 1, using heptanoylchloride (0.12 mL, 0.77 mmol). The remaining residue was purified bynormal-phase column chromatography (silica gel flash column, 0-10%MeOH/DCM), followed by reverse-phase column chromatography (RediSep PrepC18 column, 40-75% H₂O/CH₃CN). The free-base form of the desiredcompound was generated following general procedure 2, using immobilizedbicarbonate resin (0.337 g, 0.701 mmol) to afford the desired product asan amorphous white solid (0.068 g, 0.12 mmol, 45% yield). LC-MS: m/z[M+H]⁺=580.3 (calculated: 580.30); ¹H NMR: 400 MHz DMSO-d₆ δ12.04 (s,1H), 9.44 (s, 1H), 6.31 (s, 1H), 5.59 (s, 1H), 4.98 (s, 2H), 4.02 (app.s, 2H), 3.11 (q, J=7.2 Hz, 2H), 2.80 (s, 3H), 2.35 (t, J=7.2 Hz, 2H),2.19 (s, 3H), 1.96 (m, 3H), 1.85 (m, 2H), 1.69 (m, 5H), 1.53 (m, 2H),1.22 (m, 9H), 0.84 (app. m, 3H).

Example 6: cyclohexyl((2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methyl)carbonate

A solution of(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(1.0 equiv., 0.12 g, 0.26 mmol) in 2:1 DMF/pyridine (3.0 mL) was cooledon ice, and cyclohexyl chloroformate (3.0 equiv., 0.12 mL, 0.81 mmol)was added dropwise in 3 portions of 40 μL over 15 min. The mixture wasstirred on ice for 4 h. Further cyclohexyl chloroformate (3.0 equiv.,0.12 mL, 0.81 mmol) was added dropwise in 2 portions of 60 μL over 10min. The mixture was stirred on ice for 1 h. Hydrazine (15 equiv., 0.121mL, 3.85 mmol) was added, and the mixture was stirred at ambienttemperature for 35 min and concentrated. The remaining residue waspurified by reverse-phase column chromatography (RediSep Prep C18column, 40-75% H₂O/CH₃CN). The free-base form of the desired compoundwas generated following general procedure 2, using immobilizedbicarbonate resin (0.46 g, 0.96 mmol) to afford the desired product asan amorphous white solid (0.085 g, 0.124 mmol, 54% yield). LC-MS: m/z[M+H]⁺=594.3 (calculated: 594.28); ¹H NMR: 400 MHz DMSO-d₆ δ12.04 (s,1H), 9.46 (s, 1H), 6.32 (s, 1H), 5.58 (s, 1H), 5.03 (s, 2H), 4.55 (tt,J=12.6 Hz, 4.2 Hz, 1H), 4.02 (app. s, 2H), 3.12 (q, J=7.3 Hz, 2H), 2.80(s, 3H), 2.19 (s, 3H), 1.96 (m, 3H), 1.85 (m, 4H), 1.66 (m, 7H), 1.36(m, 6H), 1.21 (t, J=7.3 Hz, 3H).

Example 7:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methyltetrahydro-2H-pyran-4-carboxylate

Diisopropylethylamine (4.0 equiv., 0.209 mL, 1.20 mmol) was added to asolution of (2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(1.0 equiv., 0.14 g, 0.30 mmol), tetrahydropyran-4-carboxylic acid (2.5equiv., 0.097 g, 0.75 mmol), and N,N-dimethylaminopyridine (0.2 equiv.,7.3 mg, 0.060 mmol) in DMF (3.0 mL), and the mixture was stirred atambient temperature for 15 min. HATU (2.2 equiv., 0.25 g, 0.66 mmol) wasadded, and the mixture was stirred at ambient temperature for 16 h.Hydrazine (3 equiv., 0.028 mL, 0.90 mmol) was added, and the mixture wasstirred at ambient temperature for 30 min. Acetic acid (10 equiv., 0.17mL, 3.0 mmol) was added, and the mixture was concentrated. The remainingresidue was purified by normal-phase column chromatography (silica gelflash column, 0-10% MeOH/DCM) to afford the desired product as anamorphous white solid (0.158 g, 0.27 mmol, 89% yield). LC-MS: m/z[M+H]⁺=580.3 (calculated: 580.26); ¹H NMR: 400 MHz DMSO-d₆ δ12.04 (s,1H), 9.43 (s, 1H), 6.32 (s, 1H), 5.57 (s, 1H), 5.02 (d, J=1.9 Hz, 2H),4.02 (app. s, 2H), 3.82 (dt, J=11.4 Hz, 3.8 Hz, 2H), 3.36 (td, J=11.4Hz, 2.4 Hz, 2H), 3.12 (q, J=7.2 Hz, 2H), 2.80 (s, 3H), 2.66 (m, 1H),2.19 (s, 3H), 1.97 (m, 3H), 1.85 (m, 2H), 1.64 (m, 5H), 1.26 (app. q,J=5.9 Hz, 4H), 1.21 (t, J=7.3 Hz, 3H).

Example 8:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methylisopropyl carbonate

A solution of(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(1.0 equiv., 0.14 g, 0.30 mmol) in 2:1 DMF/pyridine (3.0 mL) was cooledon ice, and isopropyl chloroformate (1.0 M solution in toluene, 2.5equiv., 0.75 mL, 0.75 mmol) was added dropwise. The mixture was stirredon ice for 1 h. Further isopropyl chloroformate (1.0 M solution intoluene, 3.0 equiv., 0.90 mL, 0.90 mmol) was added dropwise, and themixture was stirred at ambient temperature for 16 h. Hydrazine (10equiv., 0.094 mL, 3.0 mmol) was added, and the mixture was stirred atambient temperature for 2.5 h and concentrated. The remaining residuewas purified by normal-phase column chromatography (silica gel flashcolumn, 0-10% MeOH/DCM) to afford the desired product as an amorphouswhite solid (0.094 g, 0.17 mmol, 56% yield). LC-MS: m/z [M+H]⁺=554.3(calculated: 554.25); ¹H NMR: 400 MHz DMSO-d₆ δ12.05 (s, 1H), 9.46 (s,1H), 6.33 (s, 1H), 5.57 (s, 1H), 5.02 (d, J=1.8 Hz, 2H), 4.77 (sep, 1H),4.02 (app. s, 2H), 3.12 (q, J=7.3 Hz, 2H), 2.80 (s, 3H), 2.19 (s, 3H),1.96 (m, 3H), 1.85 (m, 2H), 1.67 (m, 5H), 1.22 (d, J=6.4 Hz, 6H), 1.21(t, J=7.2 Hz, 3H).

Example 9:(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methyl(tetrahydro-2H-pyran-4-yl) carbonate

A solution of pyridine (0.63 mL, 7.8 mmol) in THF (4.0 mL) was addeddropwise to an ice-cooled solution of triphosgene (0.77 g, 2.6 mmol) inTHF (9.0 mL). The mixture was stirred on ice for 10 min. A solution oftetrahydro-4-pyranol (0.47 mL, 4.9 mmol) in THF (7.0 mL) was added, andthe mixture was stirred at ambient temperature for 1 h. The mixture wasdiluted with EtOAc (25 mL) and washed with water (35 mL), followed by0.2 M aq. HCl (25 mL) and brine (25 mL). The organic layer was driedover Na₂SO₄, filtered, and concentrated to afford a pale orange liquid,which was used without further purification in the subsequent step (0.80g, 4.9 mmol, 99% yield).

A solution of(2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-5-fluoro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(1.0 equiv., 0.14 g, 0.30 mmol) in 2:1 THF/pyridine (3.0 mL) was cooledon ice, and a solution of tetrahydro-2H-pyran-4-yl carbonochloridate(4.0 equiv., 0.20 g, 1.2 mmol) in THF (0.5 mL) was added dropwise. Themixture was stirred at ambient temperature for 1.5 h and subsequently at60° C. for 1.5 h. Further tetrahydro-2H-pyran-4-yl carbonochloridate(4.0 equiv., 0.20 g, 1.2 mmol) in THF (0.5 mL) was added, and themixture was stirred at 60° C. for 16 h. Hydrazine (24 equiv., 0.23 mL,7.2 mmol) was added, and the mixture was stirred at ambient temperaturefor 35 min. The mixture was concentrated, and the remaining residue waspurified by normal-phase column chromatography (silica gel flash column,0-5% MeOH/DCM) to afford the desired product as an amorphous white solid(0.132 g, 0.22 mmol, 73% yield). LC-MS: m/z [M+H]⁺=596.2 (calculated:596.26); ¹H NMR: 400 MHz DMSO-d₆ δ12.05 (s, 1H), 9.46 (s, 1H), 6.33 (s,1H), 5.58 (s, 1H), 5.05 (d, J=1.6 Hz, 2H), 4.77 (sep, J=6.4 Hz, 1H),4.02 (app. s, 2H), 3.77 (dt, J=11.6 Hz, 4.5 Hz, 2H), 3.44 (ddd, J=11.9Hz, 9.3 Hz, 2.9 Hz, 2H), 3.12 (q, J=7.3 Hz, 2H), 2.80 (s, 3H), 2.19 (s,3H), 1.93 (m, 7H), 1.63 (m, 7H), 1.21 (t, J=7.3 Hz, 3H).

Preparation 10: methyl2-chloro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxylate

A mixture of 5-methyl-1H-pyrazol-3-amine (5.6 g, 58 mmol), methyl2,6-dichloropyrimidine-4-carboxylate (12.0 g, 58 mmol), and DIPEA (15.0g, 116 mmol) in DMSO (120 ml) was stirred at 25° C. for 12 hours. H₂O(500 mL) was added and the precipitated solid was collected byfiltration to give the title intermediate (15 g, 97%) as a yellow solid.(m/z): [M+H]⁺ calcd for C₁₀H₁₁ClN₅O₂ 268.05 found 268.1.

Preparation 11: tert-butyl(1R,3s,5S)-3-((4-(methoxycarbonyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate

A mixture of methyl2-chloro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxylate(12.0 g, 45 mmol), tert-butyl(1R,3s,5S)-3-(methylamino)-9-azabicyclo[3.3.1]nonane-9-carboxylate (13.7g, 54 mmol), and DIPEA (12.0 g, 90 mmol) in NMP (120 ml) was stirred at120° C. for 16 hours. The reaction was poured into H₂O (2000 mL), theprecipitated solid was collected by filtration to give the titleintermediate (15 g, 68%) as a white solid. (m/z): [M+H]⁺ calcd forC₂₄H₃₆N₇O₄ 486.28 found 486.3.

Preparation 12: tert-butyl(1R,3s,5S)-3-((4-carbamoyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate

To tert-butyl(1R,3s,5S)-3-((4-(methoxycarbonyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate(3 batches of 2 g, 4.12 mmol) was added NH₃/MeOH (3 aliquots of 60 ml)in a 100 ml sealed tube, the reaction mixture was stirred at 25° C. for12 hours. The reaction mixture was concentrated in vacuum to afford thetitle intermediate (3.7 g, 64%). (m/z): [M+H]⁺ calcd for C₂₃H₃₅N₈O₃471.28 found 471.3.

Preparation 13:2-(((1R,3s,5S)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxamide

To a mixture oftert-butyl(1R,3s,5S)-3-((4-carbamoyl-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-9-azabicyclo[3.3.1]nonane-9-carboxylate(3.7 g, 7.9 mmol) in dioxane (185 mL) was added HCl/Dioxane (37 mL). Thereaction was stirred at 25° C. for 3 hours. TLC showed no startingmaterial remained. The solvent was removed, and the crude product waswashed with ethyl acetate/MeOH (100:1) to give the title intermediate asthe HCl salt (4.0 g, 95%). (m/z): [M+H]⁺ calcd for C₁₈H₂₇N₈O 371.23found 371.1.

Preparation 14:2-(((1R,3s,5S)-9-(ethylsulfonyl)-9-azabicyclo[3.3.1]nonan-3-yl)(methyl)amino)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxamide(C-1)

2-))1R,3s,5S)-9-azabicyclo[3.3.1]nonan-3-yl(methyl)amino)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxamide(40 mg, 0.108 mmol) and DIPEA (0.057 ml, 0.324 mmol) were dissolved inDMF (1.50 ml) and cooled to 0° C. Ethane sulfonyl chloride was added andthe reaction mixture was allowed to warm to room temperature and stirredfor 72 hours. The reaction mixture was concentrated in vacuo and crudeproduct was purified by preparative reverse phase HPLC (Agilent Dynamax250×21.4 mm 10 μm, 15 mL/min, 2-70% ACN+0.1% TFA/ACN) to provide the TFAsalt of the title compound (4.5 mg, 9.01%). (m/z): [M+H]⁺ calcd forC₂₀H₃₁N₈O₃S 463.22 found 463.2.

Preparation 15: tert-butyl(1R,3s,5S)-3-((4-(methoxycarbonyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-8-azabicyclo[3.2.1]octane-8-carboxylate

A mixture of methyl2-chloro-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidine-4-carboxylate(8.3 g, 31.0 mmol), tert-butyl(1R,3s,5S)-3-(methylamino)-8-azabicyclo[3.2.1]octane-8-carboxylate (8.2g, 34.1 mmol), and DIPEA (10.8 mL, 62.0 mmol) in DMSO (85 ml) wasstirred at 120° C. for 16 hours. The mixture was poured into 2 L ofwater, stirred vigorously, and then filtered to afford the titlecompound (11.1 g, 76%). (m/z): [M+H]⁺ calcd for C₂₃H₃₄N₇O₄ 472.27 found472.3.

Preparation 16: tert-butyl(1R,3s,5S)-3-((4-(hydroxymethyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-8-azabicyclo[3.2.1]octane-8-carboxylate

To a mixture of NaBH₄ (8 g, 212 mmol) in MeOH (100 mL) was addedtert-butyl(1R,3s,5S)-3-((4-(methoxycarbonyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-8-azabicyclo[3.2.1]octane-8-carboxylate(10 g, 21.2 mmol) in THF (100 mL) at 0° C. The reaction mixture was thenheated to reflux for 1 h. The reaction was quenched with water (500 mL),and the mixture extracted with ethyl acetate (3×200 mL). The combinedorganic layers were washed with brine (1×100 mL), dried over anhydrousNa₂SO₄, and concentrated in vacuo. The crude residue was purified byflash chromatography on silica gel (Petroleum ether:ethyl acetate=4:1)to afford the title compound (7 g, 68%). (m/z): [M+H]⁺ calcd forC₂₂H₃₄N₇O₃ 444.27 found 444.3.

Preparation 17:(2-(((1R,3s,5S)-8-azabicyclo[3.2.1]octan-3-yl)(methyl)amino)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol

A mixture of tert-butyl(1R,3s,5S)-3-((4-(hydroxymethyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-8-azabicyclo[3.2.1]octane-8-carboxylate(6.5 g, 14.7 mmol) in HCl/dioxane (100 mL) was stirred at r.t. for 1 h.The mixture was concentrated in vacuum to afford the HCl salt of thetitle intermediate (4.8 g, 100%). (m/z): [M+H]⁺ calcd for C₁₇H₂₆N₇O344.22 found 344.1.

Preparation 18:3-((1R,3s,5S)-3-((4-(hydroxymethyl)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-2-yl)(methyl)amino)-8-azabicyclo[3.2.1]octan-8-yl)propanenitrile (C-2)

(2-(((1R,3s,5S)-8-azabicyclo[3.2.1]octan-3-yl)(methyl)amino)-6-((5-methyl-1H-pyrazol-3-yl)amino)pyrimidin-4-yl)methanol(50 mg, 0.146 mmol) and DIPEA (0.076 ml, 0.437 mmol) were dissolved inMeOH (1.50 ml). Acrylonitrile (0.014 ml, 0.218 mmol) was added and thereaction mixture was stirred at room temperature for 90 min. Thereaction mixture was then concentrated in vacuo and the crude residuewas purified by preparative reverse phase HPLC (Agilent Dynamax 250×21.4mm 10 μm, 15 mL/min, 2-60% ACN+0.1% TFA/ACN) to provide the TFA salt ofthe title compound (14 mg, 19%). (m/z): [M+H]⁺ calcd for C₂₀H₂₉N₈O397.25 found 397.1.

Biological Assays

Assay 1: Biochemical JAK and Tyk2 Kinase Assays

A panel of four LanthaScreen JAK biochemical assays (JAK1, 2, 3 andTyk2) were carried in a common kinase reaction buffer (50 mM HEPES, pH7.5, 0.01% Brij-35, 10 mM MgCl₂, and 1 mM EGTA). Recombinant GST-taggedJAK enzymes and a GFP-tagged STAT1 peptide substrate were obtained fromLife Technologies.

Serially or discretely diluted compounds were pre-incubated with each ofthe four JAK enzymes and the substrate in white 384-well microplates(Corning) at ambient temperature for 1 h. ATP was subsequently added toinitiate the kinase reactions in 10 μL total volume, with 1% DMSO. Thefinal enzyme concentrations for JAK1, 2, 3 and Tyk2 are 4.2 nM, 0.1 nM,1 nM, and 0.25 nM respectively; the corresponding Km ATP concentrationsused are 25 μM, 3 μM, 1.6 μM, and 10 μM; while the substrateconcentration is 200 nM for all four assays. Kinase reactions wereallowed to proceed for 1 hour at ambient temperature before a 10 μLpreparation of EDTA (10 mM final concentration) and Tb-anti-pSTAT1(pTyr701) antibody (Life Technologies, 2 nM final concentration) inTR-FRET dilution buffer (Life Technologies) was added. The plates wereallowed to incubate at ambient temperature for 1 h before being read onthe EnVision reader (Perkin Elmer). Emission ratio signals (520 nm/495nm) were recorded and utilized to calculate the percent inhibitionvalues based on DMSO and background controls.

For dose-response analysis, percent inhibition data were plotted vs.compound concentrations, and IC₅₀ values were determined from a4-parameter robust fit model with the Prism software (GraphPadSoftware). Results were expressed as pIC₅₀ (negative logarithm of IC₅₀)and subsequently converted to pKi (negative logarithm of dissociationconstant, Ki) using the Cheng-Prusoff equation.

Assay 2: Inhibition of IL-2 Stimulated pSTAT5 in Tall-1 T Cells

The potency of test compounds for inhibition of interleukin-2 (IL-2)stimulated STATS phosphorylation was measured in the Tall-1 human T cellline (DSMZ) using AlphaLisa. Because IL-2 signals through JAK1/3, thisassay provides a measure of JAK1/3 cellular potency.

Phosphorylated STATS was measured via the AlphaLISA SureFire UltrapSTAT5 (Tyr694/699) kit (PerkinElmer). Human T cells from the Tall-1cell line were cultured in a 37° C., 5% CO₂ humidified incubator in RPMI(Life Technologies) supplemented with 15% Heat Inactivated Fetal BovineSerum (FBS, Life Technologies), 2 mM Glutamax (Life Technologies), 25 mMHEPES (Life Technologies) and 1× Pen/Strep (Life Technologies).Compounds were serially diluted in DMSO and dispensed acoustically toempty wells. Assay media (phenol red-free DMEM (Life Technologies)supplemented with 10% FBS (ATCC)) was dispensed (4 μL/well) and platesshaken at 900rpm for 10 mins. Cells were seeded at 45,000 cells/well inassay media (4 μL/well), and incubated at 37° C., 5% CO2 for 1 hour,followed by the addition of IL-2 (R&D Systems; final concentration 300ng/mL) in pre-warmed assay media (4 μL) for 30 minutes. After cytokinestimulation, cells were lysed with 6 ul of 3× AlphaLisa Lysis Buffer(PerkinElmer) containing 1× PhosStop and Complete tablets (Roche). Thelysate was shaken at 900 rpm for 10 minutes at room temperature (RT).Phosphorylated STAT5 was measured via the pSTAT5 AlphaLisa kit(PerkinElmer). Freshly prepared acceptor bead mixture was dispensed ontolysate (5 μL) under green filtered<100 lux light. Plates were shaken at900 rpm for 2 mins, briefly spun down, and incubated for 2 hrs at RT inthe dark. Donor beads were dispensed (5 μL) under green filtered<100 luxlight. Plates were shaken at 900 rpm for 2 minutes, briefly spun down,and incubated overnight at RT in the dark. Luminescence was measuredwith excitation at 689 nm and emission at 570 nm using an EnVision plater reader (PerkinElmer) under green filtered<100 lux light.

To determine the inhibitory potency of test compounds in response toIL-2, the average emission intensity of beads bound to pSTAT5 wasmeasured in a human T cell line. IC₅₀ values were determined fromanalysis of the inhibition curves of signal intensity versus compoundconcentration. Data are expressed as pIC₅₀ (negative decadic logarithmIC₅₀) values (mean±standard deviation).

In Vitro Assay Results

The compounds of the disclosure were tested in one or more of the assaysdescribed above.

In Table 1 below, for the JAK1, JAK 2, JAK3, and TYK2 enzyme assays, Arepresents a pK_(i) value≥10 (K_(i)≤0.1 nM), B represents a pK_(i) valuebetween 9 and 10 (K_(i) between 1 nM and 0.1 nM), C represents a pK_(i)value between 8 and 9 (K_(i) between 10 nM and 1 nM), D represents apK_(i) value between 7 and 8 (K_(i) between 100 nM and 10 nM), and Erepresents a pK_(i) value of 7 or below (K_(i) of 100 nM or above). Forthe Tall-1 Potency assay, A represents a pIC₅₀ value≥8.0, and Brepresents a pIC₅₀ value between 7.5 (included) and 8.0.

TABLE 1 Tall-1 IL2 JAK 1 JAK 2 JAK 3 Tyk 2 pSTAT5 (pKi) (pKi) (pKi)(pKi) (pIC₅₀) M A A B B A 1 B A B A A 2 B B A 3 B C A 4 B C A 5 C D A 6C B D C B 7 B A B A A 8 B C A 9 B A C B A

Assay 3: Human Liver Microsome Assay

The objective of this assay was to assess the metabolic stability oftest compounds in an in vitro human liver sub-fraction. Human livermicrosomes obtained from Bioreclamation-IVT (Baltimore, Md.) were thawedon ice and diluted into 0.1M potassium phosphate buffer pH 7.4 to yieldfinal incubation protein concentrations of 0.1 mg/mL. Test compounds (10mM) were diluted into NADPH cofactor to yield final incubationconcentrations of 0.1 μmM test compound and 1 mM NADPH. Incubations wereconducted at 37° C. temperature and test aliquots were taken at timepoints 0, 5, 8, 15, 30 and 45 minutes. Each aliquot was crashed intowater with 3% formic acid and 1 μM internal standard. The resultingsamples were injected onto an LC-MS/MS system for analysis.

For each incubation, the peak area of the analytes in each t0 aliquotwas set to 100% and the peak areas from subsequent time point aliquotswere converted to percentage of parent compound remaining relative tot0. The percentage of parent compound remaining was converted to naturallog scale and plotted versus time in minutes. A linear regressionanalysis was performed for the initial decline of the parentdisappearance profile and a formula for the best-fit line determined.The slope of the resultant line was normalized to protein concentrationin mg/mL protein or number of cells/mL and CL_(int) was calculated asfollows for liver microsomes:CL _(int)(μL·min⁻¹·mg⁻¹)=(Slope×1000)/[protein, mg/mL]

CL_(int) values from 0-8 μl/min/mg represent low clearance (i.e<30% ofhepatic blood flow in human). CL_(int) values from 9-49 μl/min/mgrepresent moderate clearance (i.e. 30-70% of hepatic blood flow inhuman) and values>50 μl/min/mg represent high hepatic clearance(i.e. >70% of hepatic blood flow in human).

Compound M exhibited a HLM Cl_(int) of 132 μL/min/mg. Compounds 7 and 9exhibited a HLM Cl_(int) over 2500 μL/min/mg.

Assay 4: Aqueous Solubility Assay

The purpose of this assay was to quantify the solubility of testcompounds in pH 4 and pH 7.4 PBS buffers. The assay required 40 μL of 10mM DMSO test compound solution per desired buffer in addition to 20 μLrequired to make a test standard. For example, to test a compound inboth buffers, 100 μL (2*40 μL+20 μL) of 10 mM DMSO compound stocksolution was required.

The standard was created by diluting 20 μL of 10 mM DMSO compound stocksolution into 180 μL of methanol and was shaken for five minutes toensure solution uniformity. The resulting solution had a concentrationof 1 mM, or 1,000 μM, of the test compound. This 1,000 μM solution wasrun on an Agilent 1260 LC-MS system by injecting 2 μL in order to obtainthe peak area. For the test solutions, 40 μL of 10 mM DMSO compoundstock solution, per PBS buffer condition, were dried down into a powderovernight. Once in powder form, 400 μL of the desired PBS buffer wasadded to the powder and allowed to shake vigorously for four hours. Themaximum theoretical concentration for this sample solution was 1,000 μM.After four hours of shaking, the samples were centrifuged for 10 minutesat 3,000 RPM before injecting 2 μL on the same Agilent 1260 LC-MS systemto obtain the peak area. Once the peak areas for the standard and thetest solution were determined, the ratio of sample area to standardarea*1,000 yielded the μM solubility of the test compound solution, witha maximum upper limit of 1,000 μM. Table 2 summarizes the resultsobtained.

In Table 2 below, A represents a value above 100, B represents a valuebetween 100 and 50 (included), C represents a value between 50 and 10and D represents a value of 10 or below.

TABLE 2 Solubility at PH 7.4 (μmol) M C 1 C 2 C 3 C 4 B 5 C 7 A 6 C 9 A8 B

Assay 5: Solubility Assay in Organic Excipients

The purpose of this assay was to quantify the solubility of testcompounds in different organic excipients such as diisopropyl adipate,medium chain triglycerides (MCT), propylene glycol, and polyethyleneglycol. The assay required 80 μL of 100 mM DMSO test compound solutionper desired excipient in addition to 40 μL required to make a teststandard. For example, to test a compound in all five excipients, 440 μL(5*80 μL+40 μL) of 100 mM DMSO compound stock solution was required.

The standard was created by diluting 40 μL of 100 mM DMSO compound stocksolution into 160 μL of methanol and was shaken for five minutes toensure solution uniformity. The resulting solution had a concentrationof 20 mM, or 20,000 μM of the test compound. This 20,000 μM solution wasrun on an Agilent 1260 LC-MS system by injecting 0.2 μL in order toobtain the peak area. For the test solutions, 80 μL of 100 mM DMSOcompound stock solution, per excipient, were dried down into a powderovernight. Once in powder form, 400 μL of the desired excipient wasadded to the powder and allowed to shake vigorously for four hours. Themaximum theoretical concentration for this sample solution was 20,000μM. After four hours of shaking, the samples were centrifuged for 10minutes at 3,000 RPM before injecting 0.2 μL on the same Agilent 1260LC-MS system to obtain the peak area. Once the peak areas for thestandard and the test solution were determined, the ratio of sample areato standard area*20,000 yielded the μM solubility of the test compoundsolution, with a maximum upper limit of 20,000 μM. Table 3 summarizesthe results obtained.

In Table 3 below, A represents a value above 10, B represents a valuebetween 5 and 10, C represents a value below 5.

TABLE 3 Diiso- Pro- propyl pylene adipate Transcutol MCT glycol PEG400(mg/mL) (mg/mL) (mg/mL) (mg/mL) (mg/mL) M C A C C C 1 A A C A B 2 A A CA A 3 A A C A A 4 A A C C A 5 A A C B A 6 A A C B B 7 A C A A 8 A C C A9 A C A A

Assay 6: In Vitro Skin S9 Metabolic Stability Assay

The objective of this assay was to assess the metabolic stability ofcompounds in human and mouse skin S9 sub-fraction. Human and mouse skinS9 sub-fractions obtained from Bioreclamation-IVT (Baltimore, Md.) werethawed on ice and diluted into 0.1M potassium phosphate buffer pH 7.4 toyield final incubation protein concentrations of 1.0 mg/mL. Testcompounds (10 mM) were diluted into phosphate buffer to yield finalincubation concentrations of 0.2 μM and 2.0 μM. Incubations wereconducted at 37° C. temperature following addition of 1 mM NADPH andtest aliquots were taken at time points 0, 5, 10, 20, 30 and 45 minutes.Each aliquot was crashed into water with 3% formic acid and 1 μMinternal standard. The resulting samples were injected onto an LC-MS/MSsystem for analysis.

For each incubation, the peak area of the analytes in each t0 aliquotwas set to 100% and the peak areas from subsequent time point aliquotswere converted to percentage of parent compound remaining relative tot0. The percentage of parent compound remaining at each time point wasconverted to natural log scale and plotted versus time in minutes todetermine a disappearance half-life. The active metabolite M was alsomonitored by LC-MS in each incubation in order to demonstratecorresponding appearance of compound M to disappearance of the parentcompound. All test compounds showed release of the active metabolite M.In Table 4, the stability of compounds was assigned as follows: +++corresponds to a half-life<15 minutes, ++ corresponds to a half-lifebetween 15 and 30 minutes, and + corresponds to a half-life>30 minutes.

TABLE 4 Half-Life (min) Compound Skin S9 0.2 μM 2 μM 7-EthoxycoumarinHuman + + 7-Ethoxycoumarin Mouse + + Benfluorex Human +++ +++ BenfluorexMouse +++ +++ 1 Human + + 1 Mouse +++ ++ 2 Human + + 2 Mouse ++ ++ 3Human + + 3 Mouse +++ +++ 4 Human ++ + 4 Mouse +++ +++ 6 Human + + 6Mouse +++ +++

Assay 7: Topical Pharmacokinetics Assay

The objective of this Study was to determine the epidermal, dermal andplasma pharmacokinetics of test compounds following 24 hours of topicalexposure to intact male rat skin. Test compounds were formulated to0.25% (w/w) in ointment as described in Table 5.

TABLE 5 Formulations of Test Compounds Formulation (ointment) Testcompound  0.25% Octylhydroxystearate    5% C8-C10 Triglyceride    5%Vaseline (Petrolatum) 79.75% N-Methylpyrrolidone   10%

Twenty-four hours prior to dosing, the hair was shaved from the back of250 g male Sprague Dawley rats exposing an area 4×5 cm of at least 20cm² (about 10% of body surface). At time zero, test compound was appliedto the back of the rats at a dose of 25 μL/cm². The skin was coveredwith an adhesive cover to prevent loss of compound to the cage orbedding. Following 0.5, 2.6 and 24 h exposure, the backs were gentlywashed with soap and water to remove non-absorbed drug and patted dry.Immediately following this washing, blood was drawn by cardiac puncturefrom the rats. The outer skin (stratum corneum) was then removed byadhesive tape stripping. Upon exposure of the epidermis a 0.5 cm punchbiopsy was taken. The epidermis and dermis were quickly separated,weighed and snap frozen. Epidermis and dermis samples were homogenizedin 1:10 (w/v) water using a Covaris ultrasonic homogenizer. Samples wereextracted in 3 volumes of acetonitrile and quantified against a standardcurve via LC-MS analysis for test compound or active metabolite M. Todetermine the extent of conversion of test compound to active metaboliteM, the sum of the AUC of test compound and active metabolite M inepidermis, dermis and plasma was determined and expressed as ‘%conversion’ (i.e. ratio of active metabolite M/test compound×100).Compounds 7 and 9 exhibited a conversion over 30% in this assay.

Assay 8: Caco-2 Permeation Assay

The Caco-2 permeation assay was used as an indication of skinpermeability. The assay measures the rate at which test compounds insolution permeate a cell monolayer (designed to mimic the tight junctionof human small intestinal monolayers).

CacoReady 24-well transwell plates were obtained from ADMEcell (Alameda,Calif.). The compounds were evaluated at a concentration of 5 μM from 10mM DMSO stock solutions in duplicate (n=2). The passive permeability ofthe compounds tested was evaluated using Caco-2 cell monolayers alongwith Verapamil (25 μM) to inhibit P-gp transport proteins in the apicalto basolateral (A-B) direction. The experiment was conducted in a 37°C., 5% CO₂ incubator. Caco-2 culture media consisted of standardfiltered DMEM, FCS 10%, L-Glutamine 1% and PenStrep 1%. Basal assayplate was prepared by adding 750 μL of transport buffer to A-B wells. ACacoReady™ plate was prepared by removing the Caco-2 media from theapical wells and replacing with fresh transport media (200 μL repeatedfor a total of 3 washes). Blank media (200 μL) was then replaced withdiluted compound for A-B wells. To begin the incubation, the basal platewas removed from the incubator and the apical section was added on topof it. Samples (40 μL) were collected from the apical and basalcompartments for time zero (t0). Samples were collected again after 120minutes (t120) from the apical and basal compartments. All samples werediluted and prepared for bioanalysis by LC-MS/MS. The permeationcoefficient (K_(p), mean A to B+Verapamil Papparent) in cm/sec wascalculated as dQ (flux)/(dt×Area×concentration).

In this assay, a compound with a K_(p) value of less than about 5×10⁻⁶cm/sec is considered to have low permeability. A compound having a K_(p)value of more than about 20×10⁻⁶ cm/sec is considered to have highpermeability.

Characterization of Compound M and Comparison Compounds

TABLE 6 Characterization of comparison compounds Caco_(verap) K_(p) HLMCl_(int) Compound # Structure 10⁻⁶ cm/sec μL/min/mg M

42.3  136 C-1

 3.55  6 C-2

 5.5   12

Comparative compounds C-1 and C-2 were disclosed by applicant in somepresentations made in April, June and August 2017 at conferences.

Compound M is characterized by a much higher permeability (Cac_(Overap)value) and human liver microsome clearance (HLM Cl_(int) value) than C-1and C-2. A higher clearance is beneficial to promote quick systemicclearance and prevent systemic exposure which may be associated withside effects. Higher permeability is beneficial for skin indications asit appears to provide for better penetration in the skin.

While the present invention has been described with reference tospecific aspects or embodiments thereof, it will be understood by thoseof ordinary skilled in the art that various changes can be made orequivalents can be substituted without departing from the true spiritand scope of the invention. Additionally, to the extent permitted byapplicable patent statutes and regulations, all publications, patentsand patent applications cited herein are hereby incorporated byreference in their entirety to the same extent as if each document hadbeen individually incorporated by reference herein.

What is claimed is:
 1. A compound of formula (I):

or a pharmaceutically-acceptable salt thereof, wherein X is —O— or abond; R is selected from the group consisting of a C₁₋₈ alkyl, a 4 to 7membered heterocyclic group, and a 3 to 8 membered cycloalkyl group,wherein the C₁₋₈ alkyl, heterocyclic and cycloalkyl groups areoptionally substituted with 1 to 3 R^(a); and each R^(a) isindependently selected from the group consisting of C₁₋₄ alkyl, CN, F,OH, C₁₋₄ alkyl-OH, and C₁₋₄ alkoxy.
 2. The compound of claim 1, or apharmaceutically acceptable salt thereof, wherein R is selected from thegroup consisting of a C₁₋₆ alkyl, a 5 to 7 membered heterocyclic group,and a 5 to 7 membered cycloalkyl group, wherein the C₁₋₆ alkyl,heterocyclic and cycloalkyl groups are optionally substituted with 1 to3 R^(a).
 3. The compound of claim 1, or a pharmaceutically acceptablesalt thereof, wherein R is selected from the group consisting of a C₁₋₆alkyl, a 5 to 6 membered heterocyclic group, and a 5 to 6 memberedcycloalkyl group, wherein the C₁₋₆ alkyl, heterocyclic and cycloalkylgroups are optionally substituted with 1 to 3 R^(a).
 4. The compound ofclaim 1, or a pharmaceutically acceptable salt thereof, wherein R isselected from the group consisting of C₁₋₆ alkyl, cyclohexyl, andtetrahydropyran, wherein the C₁₋₆ alkyl, cyclohexyl, and tetrahydropyranare optionally substituted with 1 to 2 R^(a).
 5. The compound of claim1, or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom the group consisting of unsubstituted C₁₋₆ alkyl, unsubstitutedcyclohexyl, and unsubstituted tetrahydropyran.
 6. The compound of claim1, or a pharmaceutically acceptable salt thereof, wherein R is selectedfrom the group consisting of methyl, ethyl, isopropyl, propyl, n-butyl,n-hexyl, cyclohexyl, and tetrahydropyran.
 7. The compound of claim 1, ora pharmaceutically acceptable salt thereof, wherein R is C₁₋₈ alkyloptionally substituted with 1 to 3 R^(a).
 8. The compound of claim 1, ora pharmaceutically acceptable salt thereof, wherein R is a 5 to 7membered heterocyclic group optionally substituted with 1 to 3 R^(a). 9.The compound of claim 1, or a pharmaceutically acceptable salt thereof,wherein R is a 5 to 7 membered cycloalkyl group optionally substitutedwith 1 to 3 R^(a).
 10. The compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein X is a bond.
 11. The compound of claim1, or a pharmaceutically acceptable salt thereof, wherein X is —O—. 12.A compound selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 13. A compound offormula:

or a pharmaceutically acceptable salt thereof.
 14. A compound offormula:

or a pharmaceutically acceptable salt thereof.
 15. A pharmaceuticalcomposition comprising a compound of claim 1, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically-acceptable carrier. 16.The pharmaceutical composition of claim 15, further comprising one ormore additional therapeutic agents.
 17. The pharmaceutical compositionof claim 15, wherein the pharmaceutical composition is an ointment or acream.
 18. A method of treating an inflammatory or autoimmune skindisease in a mammal, the method comprising administering the compound ofclaim 1, or a pharmaceutically acceptable salt thereof, to the mammal.19. The method of claim 18, wherein the compound, or a pharmaceuticallyacceptable salt thereof, is administered to the skin of the mammal in apharmaceutical composition comprising the compound, or apharmaceutically acceptable salt thereof, and apharmaceutically-acceptable carrier.
 20. The method of claim 18, whereinthe inflammatory or autoimmune skin disease is an inflammatory skindisease.
 21. The method of claim 20, wherein the inflammatory skindisease is atopic dermatitis.
 22. The method of claim 21, wherein theatopic dermatitis is moderate to severe atopic dermatitis.
 23. Themethod of claim 21, wherein the atopic dermatitis is mild to moderateatopic dermatitis.
 24. The method of claim 18, wherein the inflammatoryor autoimmune skin disease is an autoimmune skin disease.
 25. The methodof claim 24, wherein the autoimmune skin disease is alopecia areata. 26.The method of claim 18, wherein the inflammatory or autoimmune skindisease is selected from the group consisting of: vitiligo, prurigonodularis, lichen planus, contact dermatitis, skin manifestations ofgraft versus host disease, pemphigoid, discoid lupus, lichen sclerosus,lichen planopilaris, psoriasis, and foliculitis decalvans.